Antipicornaviral compounds and methods for their use and preparation

ABSTRACT

Picornaviral 3C protease inhibitors, obtainable by chemical synthesis, inhibit or block the biological activity of picornaviral 3C proteases. These compounds, as well as pharmaceutical compositions that contain these compounds, are suitable for treating patients or hosts infected with one or more picornaviruses. Several novel methods and intermediates can be used to prepare the novel picornaviral 3C protease inhibitors of the present invention.

Continuing data: This application is a division of application Ser. No.09/226,205, filed Jan. 7, 1999, now issued as U.S. Pat. No. 6,214,799,which is a division of application Ser. No. 08/850,398, filed May 2,1997, now issued as U.S. Pat. No. 5,856,530, which claims the benefitunder 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No.60/017,666, filed May 14, 1996.

The invention pertains to the discovery and use of new compounds thatinhibit the enzymatic activity of picornaviral 3C proteases,specifically rhinovirus proteases (RVPs), as well as retard viral growthin cell culture.

The picornaviruses are a family of tiny non-enveloped positive strandedRNA containing viruses that infect humans and other animals. Theseviruses include the human rhinoviruses, human polioviruses, humancoxsackieviruses, human echoviruses, human and bovine enteroviruses,encephalomyocarditis viruses, menigovirus, foot and mouth viruses,hepatitis A virus and others. The human rhinoviruses are a major causeof the common cold. To date, there are no effective therapies to curethe common cold, only treatments that relieve the symptoms.

One strategy that may be useful to treat picornaviral infections is byinhibiting the proteolytic 3C enzymes. These enzymes are required forthe natural maturation of the picornaviruses. They are responsible forthe autocatalytic cleavage of the genomic, large polyprotein into theessential viral proteins. Members of the 3C protease family are cysteineproteases, where the sulfhydryl group most often cleaves theglutamine-glycine amide bond. In theory, inhibition of 3C proteases canblock proteolytic cleavage of the polyprotein, which in turn can retardthe maturation and replication of the viruses by interfering with viralparticle production. Therefore, inhibiting the processing of thiscysteine protease with selective, small molecules that are specificallyrecognized, may represent an important and useful approach to treat andcure viral infections of this nature and, in particular, the commoncold.

SUMMARY OF THE INVENTION

The present invention is directed to compounds that functions aspicornaviral 3C protease inhibitors, particularly those that haveantiviral activity. It is further directed to the preparation and use ofsuch 3C protease inhibitors. The Inventors demonstrate that thecompounds of the present invention bind to rhinovirus 3C proteases andpreferably have antiviral cell culture activity. The enzymaticinhibition assays used reveal that these compounds can bindirreversibly, and the cell culture assays demonstrate that thesecompounds can possess antiviral activity.

The present invention is directed to compounds of the formula (I):

wherein

R₁ is H, F, an alkyl group, OH, SH, an O-alkyl group, or an S-alkylgroup;

R₂ and R₅ are independently selected from H,

 or an alkyl group, wherein said alkyl group is different from

 with the proviso that at least one of R₂ or R₅ must be

 and wherein, when R₂ or R₅ is

X is ═CH or ═CF and Y₁ is ═CH or ═CF

or X and Y₁ together with Q′ form a three-membered ring

in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or —CF—, and Y₁ is —CH—,—CF—, or —C(alkyl)—, where R₁₀ and R₁₁ independently are H, a halogen,or an alkyl group, or, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group,

or X is —CH₂—, —CF₂—, —CHF—, or —S—, and

Y₁ is —O—, —S—, —NR₁₂—, —C(R₁₃)(R₁₄)—, —C(O)—, —C(S)—, or —C(CR₁₃R₁₄)—

wherein R₁₂ is H or alkyl, and R₁₃ and R₁₄ independently are H, F, or analkyl group, or, together with the atoms to which they are bonded, forma cycloalkyl group or a heterocycloalkyl group;

and A₁ is C, CH, CF, S, P, Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, orP—NR₁₅R₁₆

wherein R₁₅ and R₁₆ independently are an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, or a heteroaryl group,or, together with the atom to which they are bonded, form aheterocycloalkyl group;

and D₁ is a moiety with a lone pair of electrons capable of forming ahydrogen bond;

and B₁ is H, F, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈,—NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈

wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group, or, wherein any two of R₁₇, R₁₈, and R₁₉,together with the atom(s) to which they are bonded, form aheterocycloalkyl group;

and with the provisos that when D₁ is the moiety ≡N with a lone pair ofelectrons capable of forming a hydrogen bond, B₁ does not exist; andwhen A₁ is an sp³ carbon, B₁ is not —NR₁₇R₁₈ when D₁ is the moiety—NR₂₅R₂₆ with a lone pair of electrons capable of forming a hydrogenbond, wherein R₂₅ and R₂₆ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group;

and wherein D₁—A₁—B₁ optionally forms a nitro group where A₁ is N;

and wherein, when R₂ or R₅ is

X is ═CH or ═CF and Y₂ is ═C, ═CH or ═CF,

or X and Y₂ together with Q′ form a three-membered ring

in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or —CF—, and Y₂ is —CH—,—CF—, or —C(alkyl)—, where R₁₀ and R₁₁ independently are H, a halogen,or an alkyl group, or, together with the carbon atom to which they areattached, form a cycloalkyl group or a heterocycloalkyl group,

or X is —CH₂—, —CF₂—, —CHF—, or —S—, and

Y₂ is —O—, —S—, —N(R′₁₂)—, —C(R′₁₃)(R′₁₄)—, —C(O)—, —C(S)—, or—C(CR′₁₃R′₁₄)—

wherein R′₁₂ is H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR′₁₃,—NR′₁₃R′₁₄, —C(O)—R′₁₃, —SO₂R′₁₃, or —C(S)R′₁₃, and R′₁₃ and R′₁₄,independently are H, F, or an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group or,together with the atom to which they are attached, form a cycloalkylgroup or a heterocycloalkyl group;

and wherein any combination of Y₂, A₂, B₂, and D₂ forms a cycloalkylgroup, a heterocycloalkyl group, an aryl group, or a heteroaryl group;

and A₂ is C, CH, CF, S, P, Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, orP—NR₁₅R₁₆

wherein R₁₅ and R₁₆ independently are an alkyl group, a cycloalkylgroup, a heterocycloalkyl group, an aryl group, or a heteroaryl groupor, together with the atom to which they are bonded, form aheterocycloalkyl group;

and D₂ is a moiety with a lone pair of electrons capable of forming ahydrogen bond;

and B₂ is H, F, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈,—NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈

wherein R₁₇, R₁₈, and R₁₉ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group, or, wherein any two of R₁₇, R₁₈, and R₁₉,together with the atom(s) to which they are bonded, form aheterocycloalkyl group;

R₃ and R₆ are independently H, F, or an alkyl group;

R₄ is H, OH, or a suitable organic moiety;

Z and Z₁ are independently H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂,—SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃), —PO(NR₂₁R₂₂)(NR₂₃R₂₄),—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃,

wherein R₂₁, R₂₂, R₂₃, and R₂₄ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, an acyl group, or a thioacyl group, or wherein any two of R₂₁,R₂₂, R₂₃, and R₂₄, together with the atom(s) to which they are bonded,form a heterocycloalkyl group;

or Z₁, as defined above, together with R₁, as defined above, and theatoms to which Z and R₁ are bonded, form a cycloalkyl orheterocycloalkyl group,

or Z and Z₁, both as defined above, together with the atoms to whichthey are bonded, form a cycloalkyl or heterocycloalkyl group;

and pharmaceutically acceptable prodrugs, salts, and solvates thereof;

and wherein these compounds, pharmaceutically acceptable prodrugs,salts, and solvates preferably have antipicornaviral activity with anEC₅₀ less than or equal to 100 μM in the HI-HeLa cell culture assay, andmore preferably antirhinoviral activity with an EC₅₀ less than or equalto 100 μM in the HI-HeLa cell culture assay and/or anticoxsachieviralactivity with an EC₅₀ less than or equal to 100 μM in the HI-HeLa cellculture assay.

The present invention is also directed to several methods of preparingcompounds of formula (I), defined above. One method according to theinvention involves converting a compound of formula Q

wherein R₁, R₂ and R₅ are as defined above, and P₁ is a protectivegroup, preferably benzyloxy carbonyl or t-butoxycarbonyl, or a salt orsolvate thereof, to a compound of formula I, as defined above, or apharmaceutically acceptable prodrug, salt or solvate thereof.

Another method according to the invention involves converting a compoundof the formula B:

wherein R₁, R₂ and R₅ are as defined above, or a salt or solvatethereof, to a compound of formula I, as defined above, or apharmaceutically acceptable prodrug, salt or solvate thereof.

Another method according to the invention involves converting a compoundof formula O:

wherein R₁, R₂, R₅, Z and Z₁ are as defined above and P₁ is a protectivegroup, preferably benzyloxy carbonyl or t-butoxycarbonyl, or a salt orsolvate thereof, to a compound of formula I, as defined above, or apharmaceutically acceptable prodrug, salt or solvate thereof.

Another method according to the present invention involves converting acompound of formula P:

wherein R₁, R₂, R₅, Z and Z₁ are as defined above, or a salt or solvatethereof, to a compound of formula I, as defined above, or apharmaceutically acceptable prodrug, salt or solvate thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds of the formula I

wherein R₁, R₂, R₃, R₄, R₅, R₆, Z and Z₁ are as defined above, and tothe pharmaceutically acceptable prodrugs, salts, and solvates thereof,where these compounds, pharmaceutically acceptable prodrugs, salts, andsolvates preferably have antipicornaviral activity with an EC₅₀ lessthan or equal to 100 μM in the HI-HeLa cell culture assay, and morepreferably antirhinoviral activity with an EC₅₀ less than or equal to100 μM in the HI-HeLa cell culture assay and/or anticoxsachieviralactivity with an EC₅₀ less than or equal to 100 μM in the HI-HeLa cellculture assay.

The present invention preferably relates to compounds of the formula II:

wherein

R₃₁ is H, F or an alkyl group;

R₃₂ is selected from one of the following moieties:

wherein

R₃₅ is H, an alkyl group, an aryl group, —OR₃₈, or —NR₃₈R₃₉, and

R₃₆ is H or an alkyl group,

or R₃₅ and R₃₆, together with the atom(s) to which they are attached,form a heterocycloalkyl group or a heteroaryl group;

R₄₁ is H, an alkyl group, an aryl group, —OR₃₈, —SR₃₉, —NR₃₈R₃₉,—NR₄₀NR₃₈R₃₉, or —NR₃₈OR₃₉, or R₄₁ and R₃₆, together with the atom(s) towhich they are attached, form a heterocycloalkyl group;

R₃₇ is an alkyl group, an aryl group, or —NR₃₈R₃₉;

wherein R₃₈, R₃₉, and R₄₀ independently are H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, or an acyl group, or, wherein any two of R₃₈, R₃₉, and R₄₀,together with the atom(s) to which they are bonded, form aheterocycloalkyl group;

n is 0, 1 or 2;

R₃₃ is H or an alkyl group;

R₃₄ is an alkyl group, a cycloalkyl group, a heterocycloalkyl group, anaryl group, a heteroaryl group, an O-alkyl, an O-cycloalkyl group, anO-heterocycloalkyl group, an O-aryl group, an O-heteroaryl group, anS-alkyl group, an NH-alkyl group, an NH-aryl group, an N,N-dialkylgroup, or an N,N-diaryl group; and

Z and Z₁ are independently H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂,—SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃), —PO(NR₂₁R₂₂)(NR₂₃R₂₄),—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃,

wherein R₂₁, R₂₂, R₂₃, and R₂₄ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, an acyl group, or a thioacyl group, or wherein any two of R₂₁,R₂₂, R₂₃, and R₂₄, together with the atom(s) to which they are bonded,form a heterocycloalkyl group,

or Z and Z₁, both as defined above, together with the atoms to whichthey are bonded, form a heterocyclo alkyl group;

and pharmaceutically acceptable prodrugs, salts, and solvates thereof.

As used in the present application, the following definitions apply:

An “alkyl group” is intended to mean a straight or branched chainmonovalent radical of saturated and/or unsaturated carbon atoms andhydrogen atoms, such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, ethenyl, pentenyl, butenyl, propenyl, ethynyl,butynyl, propynyl, pentynl, hexynyl, and the like, which may beunsubstituted (i.e., containing only carbon and hydrogen) or substitutedby one or more suitable substituents as defined below.

A “cycloalkyl group” is intended to means a non-aromatic, monovalentmonocyclic, bicyclic, or tricyclic radical containing 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, or 14 carbon ring atoms, each of which may besaturated or unsaturated, and which may be unsubstituted or substitutedby one or more suitable substituents as defined below, and to which maybe fused one or more heterocycloalkyl groups, aryl groups, or heteroarylgroups, which themselves may be unsubstituted or substituted by one ormore suitable substituents. Illustrative examples of cycloalkyl groupsinclude, but are not limited to, the following moieties:

A “heterocycloalkyl group” is intended to mean a non-aromatic,monovalent monocyclic, bicyclic, or tricyclic radical, which issaturated or unsaturated, containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, or 18 ring atoms, and which includes 1, 2, 3, 4, or5 heteroatoms selected from nitrogen, oxygen and sulfur, wherein theradical is unsubstituted or substituted by one or more suitablesubstituents as defined below, and to which may be fused one or morecycloalkyl groups, aryl groups, or heteroaryl groups, which themselvesmay be unsubstituted or substituted by one or more suitablesubstituents. Illustrative examples of heterocycloalkyl groups include,but are not limited to the following moieties:

An “aryl group” is intended to mean an aromatic, monovalent monocyclic,bicyclic, or tricyclic radical containing 6, 10, 14, 18 carbon ringatoms, which may be unsubstituted or substituted by one or more suitablesubstituents as defined below, and to which may be fused one or morecycloalkyl groups, heterocycloalkyl groups, or heteroaryl groups, whichthemselves may be unsubstituted or substituted by one or more suitablesubstituents. Illustrative examples of aryl groups include, but are notlimited to, the following moieties:

A “heteroaryl group” is intended to mean an aromatic monovalentmonocyclic, bicyclic, or tricyclic radical containing 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, including 1, 2, 3, 4,or 5 heteroatoms selected from nitrogen, oxygen and sulfur, which may beunsubstituted or substituted by one or more suitable substituents asdefined below, and to which may be fused one or more cycloalkyl groups,heterocycloalkyl groups, or aryl groups, which themselves may beunsubstituted or substituted by one or more suitable substituents.Illustrative examples of heteroaryl groups include, but are not limitedto, the following moieties:

An “acyl group” is intended to mean a —C(O)—R radical, wherein R is anysuitable substituent as defined below.

A “thioacyl group” is intended to mean a —C(S)—R radical, wherein R isany suitable substituent as defined below.

A “sulfonyl group” is intended to mean a —SO₂R radical, wherein R is anysuitable substituent as defined below.

The term “suitable substituent” is intended to mean any of thesubstituents recognizable, such as by routine testing, to those skilledin the art as not adversely affecting the inhibitory activity of theinventive compounds. Illustrative examples of suitable substituentsinclude, but are not limited to, hydroxy groups, oxo groups, alkylgroups, acyl groups, sulfonyl groups, mercapto groups, alkylthio groups,alkoxy groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups,heteroaryl groups, carboxy groups, amino groups, alkylamino groups,dialkylamino groups, carbamoyl groups, aryloxy groups, heteroaryloxygroups, arylthio groups, heteroarylthio groups, and the like.

The term “suitable organic moiety” is intended to mean any organicmoiety recognizable, such as by routine testing, to those skilled in theart as not adversely affecting the inhibitory activity of the inventivecompounds. Illustrative examples of suitable organic moieties include,but are not limited to, hydroxy groups, alkyl groups, oxo groups,cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroarylgroups, acyl groups, sulfonyl groups, mercapto groups, alkylthio groups,alkoxy groups, carboxy groups, amino groups, alkylamino groups,dialkylamino groups, carbamoyl groups, arylthio groups, heteroarylthiogroups, and the like.

A “hydroxy group” is intended to mean the radical —OH.

An “amino group” is intended to mean the radical —NH₂.

An “alkylamino group” is intended to mean the radical —NHR where R is analkyl group as defined above.

A “dialkylamino group” is intended to mean the radical —NR_(a)R_(b)where R_(a) and R_(b) are each independently an alkyl group as definedabove.

An “alkoxy group” is intended to mean the radical —OR where R is analkyl group as defined above, for example, methoxy, ethoxy, propoxy andthe like.

An “alkoxycarbonyl group” is intended to mean the radical —C(O)OR whereR is an alkyl group as defined above.

An “alkylsulfonyl group” is intended to mean the radical —SO₂R where Ris an alkyl group as defined above.

An “alkylaminocarbonyl group” is intended to mean the radical —C(O)NHRwhere R is an alkyl group as defined above.

A “dialkylaminocarbonyl group” is intended to mean the radical—C(O)NR_(a)R_(b) where R_(a) and R_(b) are each independently an alkylgroup as defined above.

A “mercapto group” is intended to mean the radical —SH.

An “alkylthio group” is intended to mean the radical —SR where R is analkyl group as defined above.

A “carboxy group” is intended to mean the radical —C(O)OH.

A “carbamoyl group” is intended to mean the radical —C(O)NH₂.

An “aryloxy group” is intended to mean the radical —OR_(c) where R_(c)is an aryl group as defined above.

A “heteroaryloxy group” is intended to mean the radical —OR_(d) whereR_(d) is a heteroaryl group as defined above.

An “arylthio group” is intended to mean the radical —SR_(c) where R_(c)is an aryl group as defined above.

A “heteroarylthio group” is intended to mean the radical —SR_(d) whereR_(d) is a heteroaryl group as defined above.

A “pharmaceutically acceptable prodrug” is intended to mean a compoundthat may be converted under physiological conditions or by solvolysis toa compound of formula I or formula II.

A “pharmaceutically acceptable solvate” is intended to mean a solvatethat retains the biological effectiveness and properties of thebiologically active components of compounds of formulas I and II.

Examples of pharmaceutically acceptable solvates include, but are notlimited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate,acetic acid, and ethanolamine.

A “pharmaceutically acceptable salt” is intended to a mean a salt thatretains the biological effectiveness and properties of the free acidsand bases of compounds of formulas I and II and that is not biologicallyor otherwise undesirable.

Example of pharmaceutically acceptable salts include, but are notlimited to, sulfates, pyrosulfates, bisulfates, sulfites, bisulfites,phosphates, monohydrogenphosphates, dihydrogenphosphates,metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates,propionates, decanoates, caprylates, acrylates, formates, isobutyrates,caproates, heptanoates, propiolates, oxalates, malonates, succinates,suberates, sebacates, fumarates, maleates, butyne-1,4-dioates,hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates,sulfonates, xylenesulfonates, phenylacetates, phenylpropionates,phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates,tartrates, methane-sulfonates, propanesulfonates,naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.

If the inventive compound is a base, the desired salt may be prepared byany suitable method known to the art, including treatment of the freebase with an inorganic acid, such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, or withan organic acid, such as acetic acid, maleic acid, succinic acid,mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid,glycolic acid, salicylic acid, pyranosidyl acids such as glucuronic acidand galacturonic acid, alpha-hydroxy acids such as citric acid andtartaric acid, amino acids such as aspartic acid and glutamic acid,aromatic acids such as benzoic acid and cinnamic acid, sulfonic acidssuch a p-toluenesulfonic acid or ethanesulfonic acid, or the like.

If the inventive compound is an acid, the desired salt may be preparedby any suitable method known to the art, including treatment of the freeacid with an inorganic or organic base, such as an amine (primary,secondary or tertiary), an alkali metal or alkaline earth metalhydroxide or the like. Illustrative examples of suitable salts includeorganic salts derived from amino acids such as glycine and arginine,ammonia, primary, secondary and tertiary amines, and cyclic amines suchas piperidine, morpholine and piperazine, and inorganic salts derivedfrom sodium, calcium, potassium, magnesium, manganese, iron, copper,zinc, aluminum and lithium.

In the case of compounds, salts, or solvates that are solids, it isunderstood by those skilled in the art that the inventive compounds,salts, and solvates may exist in different crystal forms, all of whichare intended to be within the scope of the present invention.

The inventive compounds may exist as single stereoisomers, racematesand/or mixtures of enantiomers and/or diastereomers. All such singlestereoisomers, racemates and mixtures thereof are intended to be withinthe scope of the present invention. Preferably, the inventive compoundsare used in optically pure form.

As generally understood by those skilled in the art, an optically purecompound is one that is enantiomerically pure. As used herein, the term“optically pure” is intended to mean a compound which comprises at leasta sufficient amount of a single enantiomer to yield a compound havingthe desired pharmacological activity. Preferably, “optically pure” isintended to mean a compound that comprises at least 90% of a singleisomer (80% enantiomeric excess), preferably at least 95% (90% e.e.),more preferably at least 97.5% (95% e.e.), and most preferably at least99% (98% e.e.).

Preferably in the above formulas I and II, R₁ and R₃₁ are H or F.Preferably in formula I, R₄ is an acyl group or a sulfonyl group.Preferably in formulas I and II, D₁ and D₂ are —OR₂₅, ═O, ═S, ≡N, ═NR₂₅,or —NR₂₅R₂₆, wherein R₂₅ and R₂₆ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group, or, together with the nitrogen atom to which they arebonded, form a heterocycloalkyl group, and more preferably D₁ and D₂ are═O. Preferably A₁ and A₂ are C, CH, S, or S(O), and more preferably A₁and A₂ are C.

Preferably B₁ and B₂ are NR₁₇R₁₈, wherein R₁₇ and R₁₈ are independentlyH, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, an acyl group, or wherein R₁₇ and R₁₈,together with the atom(s) to which they are bonded, form aheterocycloalkyl group.

Preferably Z and Z₁ are independently H, an aryl group, or a heteroarylgroup, —C(O)R₂₁, —CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁,—C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂),—SONR₂₁, —SO₃R₂₁, —C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃; wherein R₂₁,R₂₂, and R₂₃ are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup, or wherein any two of R₂₁, R₂₂, and R₂₃, together with theatom(s) to which they are bonded, form a heterocycloalkyl group, or Zand Z₁, together with the atoms to which they are attached, form aheterocycloalkyl group.

Preferably R₃₂ is one of the following moieties:

wherein R₃₅, R₃₆, R₃₇, R₄₁ and n are as defined above.

Compounds according to formula I include the following, where *indicates point of attachment:

Compounds 2, 3, 4, 5, 7, 11, 12, 13, 14, 16, 17, 18, 19, 21, 22, 24, 25,41-43, 74, and 75 having the formula III:

2. R₂ is CH₂CH₂C(O)NHCPh₃, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

3. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

4. R₂ is CH₂NHC(O)CH₃; R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

5. R₂ is

R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

7. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is CO₂CH₃, and Z₁ is H

11. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₃

12. R₂ is CH₂CH₂S(O)CH₃, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

13. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is C(O)CH₃

14. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CN

16. R₂ is CH₂NHC(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃

17. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH(CH₃)₂

18. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

19. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

21. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

22. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is C(O)N(CH₃)₂

24. R₂ is CH₂CH₂C(O)NH₂; R₁ is H, Z is H, and Z₁ is C(O)Ph

25. R₂ is CH₂CH₂C(O)NH₂; R₁ is H, Z is H, and Z₁ is

41. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; Z is H; and Z₁ is

42. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

43. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; Z is H; and Z₁ is

74. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; Z is H; and Z₁ is CH₂Cl

75. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; Z is H; and Z₁ is

Compounds (26, 27, and 28) having the formula IV:

where X₁ and X₂ independently are H, F, or Cl,

26. R₂ is CH₂CH₂C(O)NH₂, X₁ is Cl and X₂ is H

27. R₂ is CH₂CH₂C(O)NH₂, X₁ is F and X₂ is H

28. R₂ is CH₂CH₂C(O)NH₂, X₁ is H and X₂ is F

Compounds (30-34) having the formula V:

30. R₄ is PhCH₂OC(O), X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, andZ₁ is CO₂CH₂CH₃

31. R₄ is CH₃CH₂CH₂SO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H,and Z₁ is CO₂CH₂CH₃

32. R₄ is PhCH₂SO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, andZ₁ is CO₂CH₂CH₃

33. R₄ is CH₃CH₂SO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, andZ₁ is CO₂CH₂CH₃

34. R₄ is PhSO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁is CO₂CH₂CH₃

Compound 29 having the formula VI:

Compound 44 having the formula VII:

Compounds (35-37) having the formula VIII:

35. X₁ is F, R₂ is CH₂CH₂C(O)NH₂, Y is CH, Z is H, and Z₁ is CO₂CH₂CH₃

36. X₁ is H, R₂ is CH₂CH₂C(O)NH₂, Y is N, Z is H, and Z₁ is CO₂CH₂CH₃

37. X₁ is H, R₂ is CH₂CH₂C(O)NH₂, Y is CH, Z is H, and Z₁ isC(O)N(CH₃)OCH₃

Compounds 46-66 and 78 having the formula IX:

46. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

47. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆ and X₁ are H; Y is CH; Z is H;and Z₁ is

48. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

49. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

50. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

51. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

52. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is C(O)tBu

53. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ and R₆ are H; X₁ is OH; Y is CH; Zis H; and Z₁ is CO₂CH₂CH₃

54. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is C(O)C(O)CH₃

55. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is C(O)C(O)N(CH₃)₂

56. R₁ is H; R₂ is CH₂OC(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is CO₂CH₂CH₃

57. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z and Z₁ together form

where the S is preferably trans to the R₁ group

58. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; and Zand Z₁ together form

59. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is C(O)NHPh

60. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is C(O)N(CH₃)Ph

61. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

62. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅, R₆, and X₁ are H; Y is CH; Z is H;and Z₁ is

63. R₁, R₅, R₆, X₁, and Z are H; Y is CH; R₂ is CH₂CH₂C(O)NH₂; and Z₁ is

64. R₁, R₅, R₆, X₁, and Z are H; Y is CH; R₂ is CH₂CH₂C(O)NH₂; and Z₁ is

65. R₁, R₅, R₆, X₁, and Z are H; Y is CH; R₂ is CH₂CH₂C(O)NH₂; and Z₁ is

66. R₁, R₅, R₆, X₁, and Z are H; Y is CH; R₂ is CH₂CH₂C(O)NH₂; and Z₁ is

78. R₁, R₅, R₆ and X₁ are H; Y is CH; R₂ is CH₂CH₂C(O)NH₂; Z is CH₂Cl;and Z₁ is H

Compounds 67-69 having the formula X:

67. R₁, R₅, R₆, X₁, and Z are H; R₂ is CH₂CH₂C(O)NH₂; Z₁ is CO₂CH₂CH₃;and Ar is Ph

68. R₁, R₅, R₆, X₁, and Z are H; R₂ is CH₂CH₂C(O)NH₂; Z₁ is CO₂CH₃; andAr is

69. R₁, R₅, R₆, X₁, and Z are H; R₂ is CH₂CH₂C(O)NH₂; Z₁ is CO₂CH₂CH₃;and Ar is

Compounds 70-73 having the formula XI:

70. R₁, R₅, R₆, and Z are H; R₂ is CH₂CH₂C(O)NH₂; R₃ is CH₂Ph; Z₁ isCO₂CH₂CH₃; and A is

71. R₁, R₅, R₆, and Z are H; R₂ is CH₂CH₂C(O)NH₂; R₃ is CH₂Ph; Z₁ isCO₂CH₂CH₃; and A is Ph

72. R₁, R₅, R₆, and Z are H; R₂ is CH₂CH₂C(O)NH₂; A is CH₂CH(CH₃)₂; Z₁is CO₂CH₂CH₃; and R₃ is

73. R₁, R₅, R₆, and Z are H; R₂ is CH₂CH₂C(O)NH₂; A is CH₂CH(CH₃)₂; Z₁is CO₂CH₂CH₃; and R₃ is

Compounds 1, 6, 8-10, 15, 20, 23, 38-40, 76, and 77 having the formulaXII:

1. R₁ is H; R₂ is CH₂CH₂CN; R₅ is H; R₆ is H; Z is F; and Z₁ isCO₂CH₂CH₃

6. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isC(O)NHCH₂CH₃

8. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is F; and Z₁ isCO₂CH₂CH₃

9. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isSO₂CH₃

10. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isSO₂Ph

15. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isCO₂H

20. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isPO(OCH₂CH₃)₂

23. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ is

38. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ is

39. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ is

40. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ is

76. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ isCH₂OAc

77. R₁ is H; R₂ is CH₂CH₂C(O)NH₂; R₅ is H; R₆ is H; Z is H; and Z₁ is

Compound 45 having the formula XIII:

45.

Compounds 79-97, also having the formula III:

82. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is CH₃ and Z₁ is CO₂CH₂CH₃

90. R₂ is CH₂CH₂C(O)NH₂, R₁ is H, and Z and Z₁ together form

where C═O is preferably cis to the R₁ group

or wherein R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is selectedfrom:

Compounds 98-121 having formula XIV:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is CO₂CH₂CH₃and

98. R₃ is CH₂Ph and R₄ is

99. R₃ is H and R₄ is

100. R₃ is

and R₄ is

101. R₃ is CH₂Ph and R₄ is

102. R₃ is CH₂Ph and R₄ is

103. R₃ is

and R₄ is

104. R₃ is CH₂Ph and R₄ is

105. R₃ is

and R₄ is

106. R₃ is CH₂Ph and R₄ is

107. R₃ is CH₂Ph and R₄ is

108. R₃ is CH₂CH₃ and R₄ is

109. R₃ is CH₃ and R₄ is

110. R₃ is CH₂Ph and R₄ is

111. R₃ is CH₂Ph and R₄ is

112. R₃ is

and R₄ is

113. R₃ is

and R₄ is

114. R₃ is

and R₄ is

115. R₃ is CH₂Ph and R₄ is

116. R₃ is CH₂Ph and R₄ is

117. R₃ is CH₂Ph and R₄ is

118. R₃ is CH₂Ph and R₄ is

119. R₃ is CH₂Ph and R₄ is

120. R₃ is CH₂Ph and R₄ is

121. R₃ CH₂CH₂CO₂H and R₄ is

Compounds 122-130, also having the formula XIV:

wherein R₆ is H, R₁ is H, R₃ is CH₂Ph and

122. R₂ is CH₂OC(O)NHC(O)CH₂Cl, Z is H, Z₁ is CO₂CH₂CH₃ and R₄ is

123. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is CO₂CH₂CH₃ and R₄ is

124. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is

and R₄ is

125. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is NO₂, and R₄ is

126. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is

and R₄ is

127. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is

and R₄ is

128. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is

and R₄ is

129. R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is CO₂CH₂CH₃ and R₄ is

130. R₂ is CH₂CH₂C(O)NH₂, Z and Z₁ together form

and R₄ is

where C═O is preferably cis to the R₁ group.

Compounds 131-145, also having the formula XIV:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₄ is

131. R₃ is CH₂Ph, Z is H and Z₁ is

132. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

133. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

134. R₃ is CH(OH)CH₃, Z is H and Z₁ is CO₂CH₂CH₃

135. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

136. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

137. R₃ is CH₂CH₂CH₃, Z is H and Z₁ is CO₂CH₂CH₃

138. R₃ is CH₂Ph, Z is H and Z₁ is C(O)N(OH)CH₃

139. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

140. R₃ is

Z is H and Z₁ is CO₂CH₂CH₃

141. R₃ is CH₂CH(CH₃)₂, Z is H and Z₁ is CO₂CH₂CH₃

142. R₃ is CH₂SCH₃, Z is H and Z₁ is CO₂CH₂CH₃

143. R₃ is CH₂SCH₂CH₃, Z is H, and Z₁ is CO₂CH₂CH₃

144. R₃ is CH₂Ph, Z is CH₃, and Z₁ is CO₂H,

145. R₃ is CH₂Ph, Z is H, and Z₁ is

Compounds 146-155, also having the formula XIV:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(O)NH₂, Z is H, and

146. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

147. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

148. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

149. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

150. Z₁ is

R₃ is CH₂Ph, and R₄ is

151. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

152. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

153. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

154. Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

155. Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Compounds 156-173, also having formula XIV:

wherein R₆ is H, R₃ is CH₂Ph, R₂ is CH₂CH₂C(O)NH₂, and

156. R₁ is OH, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

157. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

158. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

159. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

160. R₁ is H, Z is H, Z₁ is

and R₄ is

161. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

162. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

163. R₁ is H, Z is H, Z₁ is CO₂CH₂C(CH₃)₃, and R₄ is

164. R₁ is H, Z and Z₁ together form

and R₄ is

where C═O is preferably cis to the R₁ group

165. R₁ H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

166. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

167. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃and R₄ is

168. R₁ is H, Z is CH₃, Z₁ is CO₂CH₂CH₃, and R₄ is

169. R₁ is H, Z and Z₁ together form

and R₄ is

where C═O is preferably cis to R₁

170. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

171. R₁ is H, Z is CH₃, Z₁ is CO₂CH₂CH₃, and R₄ is

172. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

173. R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

Compounds 174-188, also having the formula XIV:

wherein R₆ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, and

174. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

175. Z is CH₃, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

176. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

177. Z is CH₃, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

178. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

179. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

180. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

181. Z and Z₁ together form

R₃ is

and R₄ is

where C═O is preferably cis to the R₁ group

182. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

183. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

184. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

185. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

186. Z is H, Z₁ is CO₂CH₂Ph, R₃ is

and R₄ is

187. Z is CH₃, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph and R₄

is

188. Z is H, Z₁ is CO₂CH₂CH₂OCH₃, R₃ is

and R₄ is

189. R₃ is

R₄ is

and Z and Z₁ together form

where C═O is preferably cis to the R₁ group

190. Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Other compounds according to the invention include the followingcompounds of formula III:

wherein R₆ is H, R₁ is H, R₃ is CH₂Ph, R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁is CO₂CH₂CH₃, and R₄ is selected from the following:

wherein VAR is selected from —CH₂CH₃, —CH(CH₃)₂, —CH₂CH(CH₃)₂, —CH₂—Ph,

The present invention is further directed to methods of inhibitingpicornaviral 3C protease activity that comprises contacting the proteasefor the purpose of such inhibition with an effective amount of acompound of formula I or a pharmaceutically acceptable prodrug, salt, orsolvate thereof. For example, one can inhibit picornaviral 3C proteaseactivity in mammalian tissue by administering a compound of formula I orII or a pharmaceutically acceptable prodrug, salt, or solvate thereof.More particularly, the present invention is directed to methods ofinhibiting rhinoviral protease activity.

The activity of the inventive compounds as inhibitors of picornaviral 3Cprotease activity may be measured by any of the methods available tothose skilled in the art, including in vivo and in vitro assays.Examples of suitable assays for activity measurements include theAntiviral HI—HeLa Cell Culture Assay and Normal Human BronchialEpithelial Cell Assay, both described herein.

Administration of the compounds of the formulas I and II, or theirpharmaceutically acceptable prodrugs, salts, and solvates, may beperformed according to any of the accepted modes of administrationavailable to those skilled in the art. Illustrative examples of suitablemodes of administration include, but are not limited to, oral, nasal,parenteral, topical, transdermal and rectal.

The inventive compounds of formulas I and II, and their pharmaceuticallyacceptable prodrugs, salts, and solvates, may be administered as apharmaceutical composition in any suitable pharmaceutical formrecognizable to the skilled artisan. Suitable pharmaceutical formsinclude, but are not limited to, solid, semisolid, liquid, orlyopholized formulations, such as tablets, powders, capsules,suppositories, suspensions and aerosols. The pharmaceutical compositionmay also include suitable excipients, diluents, vehicles and carriers,as well as other pharmaceutically active agents, depending upon theintended use.

Acceptable methods of preparing suitable pharmaceutical forms of thepharmaceutical compositions are known to those skilled in the art. Forexample, pharmaceutical preparations may be prepared followingconventional techniques of the pharmaceutical chemist involving stepssuch as mixing, granulating and compressing when necessary for tabletforms, or mixing, filling and dissolving the ingredients as appropriate,to give the desired products for oral, parenteral, topical,intravaginal, intranasal, intrabronchial, intraocular, intraural and/orrectal administration.

Solid or liquid pharmaceutically acceptable carriers, diluents, vehiclesor excipients may be employed in the pharmaceutical compositions.Illustrative solid carriers include starch, lactose, calcium sulphatedihydrate, terra alba, sucrose, talc, gelatin, pectin, acacia, magnesiumstearate, and stearic acid. Illustrative liquid carriers may includesyrup, peanut oil, olive oil, saline solution, and water. The carrier ordiluent may include a suitable prolonged-release material, such asglyceryl monostearate or glyceryl distearate, alone or with a wax. Whena liquid carrier is used, the preparation may be in the form of a syrup,elixir, emulsion, soft gelatin capsule, sterile injectable liquid (e.g.solution), or a nonaqueous or aqueous liquid suspension.

A dose of the pharmaceutical composition contains at least atherapeutically effective amount of the active compound (i.e., acompound of formula I or II or a pharmaceutically acceptable prodrug,salt, or solvate thereof) and preferably is made up of one or morepharmaceutical dosage units. The selected dose may be administered to amammal, for example, a human patient, in need of treatment mediated byinhibition of 3C protease activity, by any known method of administeringthe dose including topical, for example, as an ointment or cream;orally, rectally, for example, as a suppository; parenterally byinjection; or continuously by intravaginal, intranasal, intrabronchial,intraaural or intraocular infusion.

A “therapeutically effective amount” is intended to mean that amount ofa compound of formula I or II that, when administered to a mammal inneed thereof, is sufficient to effect treatment for disease conditionsalleviated by the inhibition of the activity of one or more picarnoviral3C proteases, such as human rhinoviruses, human poliovirus, humancoxsackieviruses, encephalomyocarditis viruses, menigovirus, andhepatitis A virus. The amount of a given compound of formula I or IIthat will correspond to a “therapeutically effective amount” will varydepending upon factors such as the particular compound, the diseasecondition and the severity thereof, the identity of the mammal in needthereof, but can nevertheless be readily determined by one of skill inthe art.

“Treating” or “treatment” is intended to mean at least the mitigation ofa disease condition in a mammal, such as a human, that is alleviated bythe inhibition of the activity of one or more picarnoviral 3C proteases,such as human rhinoviruses, human poliovirus, human coxsackieviruses,encephaiomyocarditis viruses, menigovirus, and hepatitis A virus, andincludes:

(a) prophylactic treatment in a mammal, particularly when the mammal isfound to be predisposed to having the disease condition but not yetdiagnosed as having it;

(b) inhibiting the disease condition; and/or

(c) alleviating, in whole or in part, the disease condition.

The inventive compounds, and their salts, solvates, and prodrugs, may beprepared by employing the techniques available in the art using startingmaterials that are readily available. Certain novel and exemplarymethods of preparing the inventive compounds are described below.

Preferably, the inventive compounds of formulas I and II are prepared bythe novel methods of the present invention, including the four generalmethods shown below. In each of these general methods, R₁, R₂, R₃, R₄,R₅, R₆, Z, and Z₁ are as defined above.

General Method I:

In General Method I, protected amino acid A, where P₁ is an appropriateprotecting group for nitrogen, is subjected to an amide forming reactionwith amino alcohol (or salt thereof) B to produce amide C. Amide C isthen deprotected to give free amine (or salt thereof) D. Amine D andCompound E, where “Lv” is an appropriate leaving group, are subjected toa bond forming reaction generating compound F. Compound F is oxidized tointermediate G, which is then transformed into unsaturated product H. Ifprotecting groups are used on any R groups (R₁-R₆) and/or on Z and/orZ₁, product H is deprotected and/or further modified to yield“deprotected or modified H.”

An alternative method to prepare intermediate F is described as follows:

Compound E and amino acid (or salt thereof) I, where P₂ is anappropriate protecting group for oxygen, are subjected to a bond formingreaction to produce intermediate J. Intermediate J is deprotected toyield free carboxylic acid K, which is subsequently subjected to anamide forming reaction with amino alcohol (or salt thereof) B togenerate intermediate F.

Amino alcohol B can be prepared as follows:

Amino acid L, where P₁ is an appropriate protecting group for nitrogen,is converted to carbonyl derivative M, where “Lv” is a leaving group.Compound M is subjected to a reaction where “Lv” is reduced to protectedamino alcohol Q. Amino alcohol Q is deprotected to give amino alcohol B.

General Method II:

In General Method II, amino acid L, where P₁ is an appropriateprotecting group for nitrogen, is converted to a carbonyl derivative M,where “Lv” is a leaving group. Compound M is subjected to a reactionwhere “Lv” is replaced by R₁ to give derivative N. Derivative N is thentransformed into unsaturated product O. Unsaturated compound O isdeprotected to give free amine (or salt thereof) P, or modified one ormore times at R₂, R₅, Z and/or Z₁ to give one or more modified Ocompounds.

Modified O is then deprotected to give amine (or salt thereof) P. AmineP is subsequently subjected to an amide forming reaction with carboxylicacid K, prepared as described in General Method I, to give final productH. If protecting groups were used on any R group (R₁-R₆) and/or on Zand/or Z₁, product H is deprotected and/or further modified to yield“deprotected or modified H.”

An alternative method to prepare intermediate N is described as follows:

Compound M is subjected to a reaction where “Lv” is reduced to protectedamino alcohol Q. Amino alcohol Q is subsequently oxidized to derivativeN.

General Method III:

In General Method III, amino acid L, where P₁ is an appropriateprotecting group for nitrogen, is converted to a carbonyl derivative M,where “Lv” is a leaving group. Derivative M is deprotected to give freeamine (or salt thereof) R, which subsequently is subjected to an amideforming reaction with carboxylic acid K to give intermediate S.Intermediate S is then either converted directly to carbonylintermediate G, or successively reduced to alcohol F, which is thenoxidized to G. Intermediate G is subjected to a reaction to yield theunsaturated final product H. If protecting groups were used on any Rgroups (R₁-R₆) and/or on Z and/or Z₁, product H is deprotected and/orfurther modified to yield “deprotected or modified H.”

General Method IV:

In General Method IV, free amine (or salt thereof) P, prepared fromintermediate O as described in General Method II, is converted to amideT by reaction with amino acid A, where P₁ is an appropriate protectinggroup for nitrogen. Compound T is further deprotected to free amine (orsalt thereof) U, which is subsequently converted to H with reactiveintermediate E. If protecting groups were used on any R groups (R₁-R₆)and/or on Z and/or Z₁, product H is deprotected and/or further modifiedto yield “deprotected or modified H.”

Preferably the compound of formulas I or II can be prepared by one offour specific methods. For example, compounds 4, 12, 14, 16, 20, 23, 24,26-30, 35, and 36 can be prepared by Specific Method I:

In Specific Method I, carboxylic acid K, CBZ-L-Leu-L-Phe, which can bepurchased from Bachem or prepared as described in General Method I, issubjected to an amide forming reaction with amino alcohol (or saltthereof) B to generate intermediate F. Intermediate F is oxidized tointermediate G, which is then transformed into unsaturated product H. Inthe case of Compound 12, intermediate F is oxidized to modified F, whichis then oxidized to intermediate G. If protecting groups were used onany R groups (R₁-R₆) and/or on Z and/or Z₁, product H is deprotectedand/or further modified to yield “deprotected or modified H.”

For example, compounds 1-3, 6-11, 17-19, 21, 22, 25, 37-40, and 74-77can be prepared by Specific Method II:

In Specific Method II, intermediate P (or salt thereof), prepared asdescribed in General Method II, is subjected to an amide formingreaction with carboxylic acid K, CBZ-L-Leu-L-Phe, which can be purchasedfrom Bachem or prepared as described in General Method I, to give finalproduct H. If protecting groups were used on any R group (R₁-R₆) and/oron Z and/or Z₁, product H is deprotected and/or further modified toyield “deprotected or modified H.”

For example, compounds 5, 13, and 15 can be prepared by Specific MethodIII.

In Specific Method III, free amine (or salt thereof) R, prepared asdescribed in General Method III, is subjected to an amide formingreaction with carboxylic acid K, CBZ-L-Leu-L-Phe, which can be purchasedfrom Bachem or prepared as described in General Method I, to giveintermediate S. Intermediate S is then either converted directly tocarbonyl intermediate G, in the case of compounds 13 and 15, or reducedto alcohol F, which is then oxidized to intermediate G, in the case ofcompound 5. Intermediate G is subjected to a reaction to yield theunsaturated final product H. If protecting groups were used on any Rgroups (R₁-R₆) and/or on Z, and/or Z₁, product H is deprotected and/orfurther modified to yield “deprotected or modified H.”

For example, compounds 31-34 can be prepared by Specific Method IV:

In Specific Method IV, free amine (or salt thereof) P, prepared asdescribed in General Method II, is converted to amide T by reaction withprotected amino acid A, which can be purchased from Bachem, AdvancedChemtech, and Synthetech. Compound T is further deprotected to freeamine (or salt thereof) U, which is subsequently converted to H withreactive intermediate E. If protecting groups were used on any R groups(R₁-R₆) and/or on Z and/or Z₁, product H is deprotected and/or furthermodified to yield “deprotected or modified H.”

Suitable protecting groups for nitrogen are recognizable to thoseskilled in the art and include, but are not limited tobenzyloxycarbonyl, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl,p-methoxybenxyloxycarbonyl, trifluoroacetamie, and p-toluenesulfonyl.Suitable protecting groups for oxygen are recognizable to those skilledin the art and include, but are not limited to —CH₃, —CH₂CH₃, tBu,—CH₂Ph, —CH₂CH═CH₂, —CH₂OCH₂CH₂Si(CH₃)₃, and —CH₂CCl₃. Other examples ofsuitable protecting groups for nitrogen or oxygen can be found in T.Green & P. Wuts, Protecting Groups in Organic Synthesis (2nd ed. 1991),which is incorporated herein by reference.

Suitable leaving groups are recognizable to those skilled in the art andinclude, but are not limited to, Cl, Br, I, sulfonates, O-alkyl groups,

Other examples of suitable leaving groups are described in J. March,Advanced Organic Chemistry, Reactions, Mechanisms, and Structure (4thed. 1992) at pages 205, 351-56, 642-43, 647, 652-53, 666, 501, 520-21,569, 579-80, 992-94, 999-1000, 1005, and 1008, which are incorporatedherein by reference.

EXAMPLES

Examples of the processes used to make several of the compounds offormulas I and II are set forth below. The structures of the compoundsof the following Examples were confirmed by one or more of thefollowing: proton magnetic resonance spectroscopy, infraredspectroscopy, elemental microanalysis, mass spectrometry, thin layerchromatography and melting point.

Proton magnetic resonance (NMR) spectra were determined using a Tech-Magor Varian UNITYplus 300 spectrometer operating at a field strength of300 megahertz (MHz). Chemical shifts are reported in parts per million(δ) and setting the references such that in CDCl₃ the CHCl₃ is at 7.26ppm, in acetone-d₆ the acetone is at 2.02 ppm, and in DMSO-d₆ the DMSOis at 2.49 ppm. Peak multiplicities are designated as follows: s,single, d, doublet; dd, doublet of doublets; ddd, doublet of doublet ofdoublets; t, triplet; q, quartet; bs, broad single; bt, broad triplet;m, multiplet. Mass spectra (GAB; fast atom bombardment) were determinedat the Scripps Research Institute Mass Spectrometry Facility, San Diego,Calif. Infrared absorption (IR) spectra were taken on a MIDACCorporation FTIR or a Perkin-Elmer 1600 series FTIR spectrometer.

Elemental microanalysis were performed by Atlantic Microlab Inc.Norcross, Ga, and gave results for the elements stated with ±0.4% of thetheoretical values. Flash chromatography was performed using Silica gel60 (Merck At 9385). Thin layer chromatographs (TLC) were performed onprecoated sheets of silica 60 F₂₅₄ (Merck Art 5719). Melting points weredetermined on a Mel-Temp apparatus and are uncorrected. AnhydrousN,N-Dimethylformamide (DMF), N,N-dimethylacetamide (DMA),dimethylsulfoxide (DMSO), were used as is. Tetrahydrofuran (THF) wasdistilled from sodium benzophenone ketyl under nitrogen.

Et₂O refers to diethyl ether. Pet, ether refers to petroleum etherhaving a boiling range of 36-53° C. TFA refers to trifluoroacetic acid.Et₃N refers to triethylamine. Other abbreviation include; methanol(MeOH), ethanol (EtOH), ethyl acetate (EtOc), acetyl (Ac), methyl (Me),phenyl (Phe), triphenylmethyl (Tr), benzyloxycarbonyl (CBZ),tert-butoxycarbonyl (BOC), m-chloroperoxybenzoic acid (m-CPBA), alanine(Ala), glutamine (Gln), leucine (Leu), methionine (Met), phenylalanine(Phe), penicillamine (Pen). Additionally, “L” represents natural aminoacids, “D” represent unnatural amino acids, and “DL” represents racemicmixtures.

A simplified naming system was used to identify intermediates and finalproducts. Amino acid and peptide alcohols are given the suffix ‘ol’ (forexample methioninol). Amino acid and peptide aldehydes are given thesuffix ‘al’ (for example methioninal). When naming final products,italicized amino acid abbreviations represents modifications at theC-terminus of that residue where the following apply:

1. acrylic acid esters are reported as either “E” (trans) or “Z” (cis)propenoates.

2. acrylonitriles are reported as either E and Z propenonitriles,

3. acrylamides are reported as either E or Z propenamides, except in thecase of the compounds 21, which is reported as1-Pyrrolidin-1-yl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenone,

4. vinyl sulfones, vinyl phosphonates, or vinyl aryls are reported as Eor Z vinyl sulfones, vinyl phosphonates or aryls, and

5. vinyl ketones are reported as either E or Z en-2-ones.

Example 1 Preparation of Compound 12;Ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)]-E-propenoate Preparation ofIntermediate CBZ-L-Leu-L-Phe-L-Methioninol

CBZ-L-Leu-L-Phe-(3.02 g, 7.3 mmol) was dissolved in 75 ml, of CH₂Cl₂. Tothis solution was added N-hydroxysuccinimide (0.91 g, 7.7 mmol) and 2 mLof DMF, and stirring was continued until all solids had gone intosolution. N,N′-Dicyclohexylcarbodiimide (1.60 g, 7.7 mmol) was added tothe reaction mixture, and the reaction was stirred at room temperaturefor one hour. The mixture was then filtered into a separate flaskcontaining S-(−)-methioninol (1.06 g, 7.7 mmol) dissolved in a minimumof DMF, removing the N,N′-dicyclohexylurea precipitate. The reaction wasallowed to stir overnight at room temperature. The solvents were removedunder vacuum, and the resulting crude product was purified by flashchromatography (anhydrous NH₃/MeOH/CHCl₃, 0.5:4.5:9.5) on silica gel togive 3.72 g (96%) of white solid: IR (KBr) 3293, 3065, 2955, 1696, 1645,1539, 1236, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ0.80 (m, 6 H), 1.31 (m, 2 H),1.51 (m, 2 H), 1.82 (m, 1 H), 2.00 (s, 3 H), 2.43 (m, 2 H), 2.78-3.29(m, 4 H), 3.72 (m, 1H), 3.97 (m, 1 H), 4.45 (m, 1 H), 4.66 (t, 1 H,J=5.5 Hz), 5.01 (s, 2 H), 7.15-7.39 (m, 10 H), 7.43 (d, 1H, J=8.1 Hz),7.62 (d, 1 H, J=8.5 Hz), 7.95 (d, 1 H, J=8.1 Hz), Anal. (C₂₈H₃₉N₃O₃S) C,H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-Methioninol (sulfoxide)

CBZ-L-Leu-L-Phe-L-methioninol (1.50 g, 2.80 mmol) was dissolved in 50 mLof CH₂Cl₂. A total amount of 0.61 g (3.5 mmol) of m-CPBA was addedportionwise over a period of five hours as the reaction was stirred atroom temperature. After an additional hour, the reaction was poured intosaturated NaHCO₃/CH₂Cl₂. The organic layer was separated, washed withbrine, and dried (Na₂SO₄). After removal of the solvent, the cruderesidue was flash chromatographed on a short flash silica gel columneluting with 5% MeOH/CHCl₃. The product was obtained as a white glassysolid (1.38 g, 90%); IR (KBr) 3295, 3063, 2955, 1694, 1644, 1541, 1263,1234, 1043, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ0.81 (m, 6 H), 1.32 (m, 2 H),1.92 (m, 1H), 2.47 (s, 3 H), 2.55-3.29 (m, 6 H), 3,73 (m, 1 H), 3.97 (m,1 H), 4.75 (t, 1 H, J=5.5 Hz), 5.01 (m, 2 H), 7.16-7.39 (m, 10 H), 7.44(d, 1 H, J=7.7 Hz), 7.73 (d, 1 H, J=8.8Hz), 7.98 (m, 1 H), Anal.(C₂₈H₃₉N₃O₆S) C, H, N, S.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-methioninal (sulfoxide)

CBZ-L-Leu-L-Phe-L-methioninol (sulfoxide) (1.38 g, 2.53 mmol) wasdissolved in DMSO, o-lodoxybenzoic acid (2.12 g, 7.59 mmol) was added,requiring a few minutes of stirring at room temperature to dissolve.After three hours, the DMSO was removed under reduced pressure. Theresidue was twice diluted with CH₂Cl₂, and the solvent was evaporated toremove any residual DMSO. The residue was diluted with a minimum ofacetone, and the white precipitate was filtered off. The filtrate wasconcentrated to near dryness and dissolved in EtOAc, which produced moreof the white precipitate, which was again filtered off. The filtrate waswashed with a 10% Na, S₂O₃/10% NaHCO, solution, water, and brine beforedrying over Na₃SO₄. Upon removal of the organic solvent, the residue wastwice taken up in benzene and evaporated to remove any residual water,giving 0.98 g (71%) of a white glassy solid which was used immediatelywithout further purification: ¹H NMR (DMSO-d₆) δ0.81 (m, 6H), 1.30 (m,2H), 1.50 (m, 1H), 1.97 (m, 1H), 2.48 (s, 3H), 2.55-3.27 (m, 5H), 3.70(m, 1), 4.47 (m, 1H), 4.71 (m, 1H), 5.00 (s, 2H), 7.20-7.40 (m, 10H),7.93 (m, 1H), 8.08 (m, 1H), 8.51 (m, 1H), 9.22 (s, 1H); (M+H) 544.

Preparation ofProduct—Ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)]-E-Propenoate

CBZ-L-Leu-L-Phe-L-Methioninal (sulfoxide) (0.98 g, 1.80 mmol) wasdissolved in 50 mL of THF, (Carbethoxymethylene)triphenyl-phosphorane(1.11 g, 2.16 mmol) was added, and the reaction was stirred at roomtemperature overnight. The solvent was removed in vacuo, and the residuesubjected to flash column chromatography eluting with 2% MeOH/CHCl₃. Theproduct was obtained (0.82 g, 74%), as a white solid; ¹H NMR (DMSO-d₆)δ0.81 (m, 6H), 1.21 (t, 3H, J=7 Hz), 1.34 (m, 2H), 1.54 (m, 1H), 1.78(m, 1H), 1.93 (m, 1H), 2.49 (s, 3H), 2.50-3.05 (m, 4H), 3.99 (m, 1H),4.10 (q, 2H, J=7 Hz), 4.51 (m, 2H), 5.00 (dd, 2H, J=17.3, 4.4Hz), 5.62(m, 1H), 6.72 (m, 1H), 7.19 (m, 5H), 7.34 (m, 5H), 7.43 (d, 1H,J=8.1Hz), 8.08 (d, 1H, J=7.4 Hz), 8.13 (d, 1H, J=8.5Hz); (M+H) 614; HRMScalcd for C₃₂H₄₃N₃O₇S+Cs 746.1876 (M+Cs), found 746.1850, Anal.(C₃₂H₄₁N₃O₂S) C, H, N, S.

Example 2 Preparation of Compound 4;Ethyl-3-[CBZ-L-Leu-L-Phe-L-(N-Ac-amino)-Ala]-E-Propenoate Preparation ofIntermediate CBZ-L-(N-Ac-amino)-Ala

CBZ-L-Amino-Ala (1.5 g, 6.3 mmol) was suspended in 50 mL of H₂O withstirring Acetic anhydride (5.0 mL) was added slowly to this suspensionover a 30 minute period, during which time the starting materialdissolved. The reaction mixture was stirred for an additional 1 hour atroom temperature and then evaporated to dryness under vacuum. Theresulting oil was dissolved in 30 mL CHCl₃ and left for 12 hours. Thesolid that formed was collected by filtration, washed with 30 mL ofCHCl₃ and dried yielding 1.29 g (73%) of product as a white solid; IR(KBr) 3271, 3125, 3065, 1734, 1703, 1614, 1545, 1289, 1244, 1053, 727cm⁻¹; ¹H NMR (DMSO-d₆) δ1.84 (s, 3H), 3.2-3.55 (m, 2H), 4.13 (m, 1H),5.08 (s, 2H), 7.12-7.41 (m, 5H), 7.54 (d, 1H, J=8.1 Hz), 8.02 (bt, 1H,J=5.5 Hz), 12.78 (bs, 1H); Anal. (C₁₃H₁₆N₂O₅) C, H, N.

Preparation of Intermediate CBZ-L-(N-Ac-amino)-Ala-OMe

Anhydrous HCl gas was slowly bubbled at 0° C. into a stirred suspensionof CBZ-L-(N-Ac-Amino)-Ala(1.21 g, 4.3 mmol) in MeOH (43 mL) until thesolid was dissolved. Stirring was continued for 30 minutes at 0° C.whereupon the methanolic HCl was carefully evaporated to dryness. Themethyl ester was formed as a white solid in quantitative yield and usedwithout further purification: IR (KBr) 3323, 3285, 3094, 2957, 1755,1736, 1686, 1651, 1531, 1277, 1057, 736, 600 cm⁻¹; ¹H NMR (DMSO-d₆)δ1.78 (s, 3H), 3.22-3.47 (m, 2H), 3.61 (s, 3H), 4.15 (m, 1H), 5.02 (s,2H), 7.24-7.36 (m, 5H), 7.64 (d, 1H, J=7.7 Hz), 7.97 (bt, 1H, J=6.3 Hz);Anal. (C₁₄H₁₈N₂O₅) C, H, N.

Preparation of Intermediate CBZ-L-(N-Ac-amino)-Alaninol

To a solution of CBZ-L-(N-Ac-amino)-Ala-OMe (1.8 g, 6.12 mmol) in 50 mLanhydrous THF/EtOH (2:1) was added LiCl (0.52 g, 12.24 mmol). Upondissolution, NaBH₄ (0.46 g, 12.24 mmol) was added, and the mixture wasstirred at room temperature for 12 hours. The reaction mixture wasevaporated to near dryness, whereupon 45 mL of H₂O was added. The pH ofthis mixture was adjusted to 2-3 using concentrated HCl, followed byextraction with EtOAc (300 mL). The organic layer was washed with H₂O(50 mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by flash column chromatography (10% MeOH/CHCl₃) to give 1.38 g(85%) of a white solid: IR (KBr) 3303, 3082, 2951, 2926, 1689, 1645,1547, 1284, 1061, 0146, 756, 698 cm⁻¹, ¹H NMR (DMSO-d₆) δ1.78 (s, 3H),3.03 (m, 1H), 3.16-3.28 (m, 3H), 3.49 (m, 1H), 5.00 (s, 2H), 6.95 (d,1H, J=8.1 Hz), 7.29-7.38 (m, 5H), 7.83 (bt, 1H, J=5.5 Hz); Anal.(C₁₃H₁₈N₂O₄) C, H, N.

Preparation of Intermediate L-(N-Ac-amino)-Alaninol

To a solution of CBZ-L-(N-Ac-amino)-alaninol (1.36 g, 5.11 mmol) in 40mL MeOH, 10% Pd on carbon (0.15 g) was added with stirring while underan argon atmosphere. The reaction vessel was evacuated under vacuum andthen put under an atmosphere of hydrogen using a balloon. The mixturewas stirred for 2 hours. At this time the hydrogen gas was evaluated,and the catalyst was removed by filtration. The solvent was removedunder vacuum. Addition of EtOAc and reconcentration gave a whitehygroscopic solid in quantitative yield which was used without furtherpurification: mp =80-82° C.; ¹H NMR (DMSO-d₆) δ1.79 (s, 3H), 2.66 (m,1H), 2.86 (m, 1H), 3.06 (m, 1H), 3.21 (2H, m), 3.4 (bs, 2H), 4.55 (bs,1H), 7.76 (bs, 1H), Anal. (C₅H₁₂N₂O₃) C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(N-Ac-amino)-Alaninol

This compound was prepared from CBZ-L-Leu-L-Phe andL-(N-Ac-amino)-alaninol using the procedure described in Example 1 forthe preparation of CBZ-L-Leu-L-Phe-L-methioninol. The compound waspurified by column chromatography (7% MeOH/CHCl₃) to give a white solid(81%); IR (KBr) 3302, 2955, 1694, 1651, 1539, 1454, 1236, 1047, 698cm⁻¹; ¹H NMR (DMSO-d₆) δ0.80 (s, 6H), 1.32 (m, 2H), 1.47 (m, 1H), 1.79(s, 3H), 2.81 (m, 1H), 2.97 (m, 2H); 3.14-3.25 (m, 3H), 3.71 (m, 1H),3.95 (m, 1H), 4.42 (m, 1H), 4.67 (t, 1H, J=5.5 Hz), 5.00 (m, 2H),7.16-7.34 (m, 10H), 7.45 (d, 1H, J=8.1 Hz), 7.70 (m, 2H), 7.88 (d, 1H,J=8.1 Hz); Anal. (C₂₈H₃₈N₄O₄) C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(N-Ac-amino)-Alaninal

This compound was prepared in 73% yield as a white solid fromCBZ-L-Leu-L-Phe-L-(N-Ac-amino)-alaninol using the procedure described inExample 1 for the preparation of CBZ-L-Leu-L-Phe-L-methioninal(sulfoxide). The product was used immediately without furtherpurification. The product existed as a mixture of aldehyde and aldehydehydrate. IR (KBr) 3294, 2957, 1695, 1649, 1539, 1263, 698 cm⁻¹; ¹H NMR(DMSO-d₆) δ0.81 (dd, 6H, J=8.8, 6.2 Hz), 1.31 (m, 2H), 1.50 (m, 1H),1.76 (s, hydrate), 1.78 (s, hydrate), 1.78 (s, 3H), 2.83 (m, 1H), 3.00(m, 1H), 3.20 (d, J=9.6 Hz, hydrate), 3.35 (m, 1H), 3.80 (m, hydrate),3.97 (m, 2H), 4.16 (m, 1H), 4.37 (m, hydrate) 4.44 (m, hydrate), 4.54(m, 1H), 5.01 (s, 2H), 6.28 (d, 1H, J=7.0 Hz, hydrate), 6.41 (d, 1H,J=6.6 Hz, hydrate), 7.12-7.50 (m, 10H), 7.63 (t, 1H, J=7.9 Hz), 7.87 (m,1H), 7.98 (d, 1H, J=8.1 Hz), 8.40 (d, 1H, J=7.0 Hz), 9.26 (s, 1H); Anal.(C₂₈H₃₆N₄O₆·0.5 H₂O) C, H, N.

Preparation ofProduct—Ethyl-3-[CBZ-L-Leu-L-Phe-L-(N-Ac-amino)-Ala]-E-Propenoate

This compound was prepared in 55% yield as a white solid fromCBZ-L-Leu-L-Phe-L-(N-Ac-amino)-alaninal and(carbethoxymethylene)triphenylphosphorane using the procedure describedin Example 1 for the preparation of compound 12,ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate. The product waspurified by flash column chromatography (3% MeOH/CHCl₃), ¹H NMR(DMSO-d₆) δ0.81 (dd, 6H, J=9.2, 6.6 Hz), 1.21 (t, 3H, J=7.2 Hz), 1.34(m, 2H), 1.53 (m, 1H), 1.78 (s, 3H), 2.80-3.28 (m, 4H), 3.99 (m, 1H),4.10 (q, 2H, J=7.0 Hz), 4.43 (m, 2H), 5.01 (m, 2H), 5.61 (d, 1H, J=15.4Hz), 6.61 (dd, 1H, J=15.4, 5.2 Hz), 7.10-7.34 (m, 10H), 7.44 (d, 1H,J=7.7 Hz), 7.70 (m, 2H), 7.82 (t, 1H, J=5.5 Hz), 8.05 (m, 2H); HRMScalcd for C₃₂H₄₂N₄O₇+Cs 727.2108 (M+Cs), found 727.2137, Anal.(C₃₂H₄₂N₆O₇) C, H, N.

Example 3 Preparation of Compound 2;Ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate Preparation ofIntermediate BOC-L-(Tr-Gln)-N-Me)OMe

Isobutyl chloroformate (0.611 mL, 4.71 mmol) was added to a solution ofBOC-L-(Tr-Gln) (2.30 g, 4.71 mmol) and 4-methylmorpholine (1.04 mL, 9.46mmol) in CH₂Cl₂ at 0° C. The reaction mixture was stirred at 0° C. for20 minutes then N,O-dimethylhydroxylamine hydrochloride (0.459 g, 4.71mmol) was added. The resulting solution was stirred at 0° C. for 15minutes and at 23° C. for 4 hours, then was partitioned between water(150 mL) and a 1:1 mixture o EtOAc and hexanes (2×150 mL). The combinedorganic layers were dried over Na₂SO₄ and were concentrated.Purification of the residue by flash column chromatography (40% hexanesin EtOAc) afforded the product (2.22 g, 89%) as a white foam: R_(f)=0.22(50% EtOAc in hexanes); IR (KBr) 3411, 3329, 3062, 1701, 1659 cm⁻¹; ¹HNMR (CDCl₃) δ1.42 (s, 9H), 1.63-1.77 (m, 1H), 2.06-2.17 (m, 1H),2.29-2.43 (m, 2H), 3.17 (s, 3H), 3.64 (s, 3H), 4.73 (bs, 1H), 5.38-5.41(m, 1H), 7.20-7.31 (m, 15H); Anal. (C₃₁H₃₇N₃O₅) C, H, N.

Preparation of Intermediate BOC-L-(Tr-Glutaminal)

Diisobutylamuminum hydride (7.84 mL of 1.5 M solution in toluene, 11.76mmol) was added to a solution of BOC-L-(Tr-Gln)-N(Me)OMe (2.50 g, 4.70mmol) in THF at −78° C. and the reaction mixture was stirred at −78° C.for 4 hours. Methanol (3 mL) and 1.0 M HCl (6 mL) were addedsequentially, and the mixture was warmed to 23° C. The resultingsuspension was diluted with Et₂O (150 mL) and was washed with 1.0 M HCl(3×100 mL), half-saturated NaHCO₁ (100 mL), and water (100 mL). Theorganic layer was dried over MgSO₄, filtered, and concentrated to givecrude aldehyde (2.01 g, 91%) as a white solid: mp=114-116° C.;R_(f)=0.42 (50% EtOAc in hexanes); IR (KBr) 3313, 1697, 1494 cm⁻¹; ¹HNMR (CDCl₃) δ1.44 (s, 9H), 1.65-1.75 (m, 1H), 2.17-2.23 (m, 1H),2.31-2.54 (m, 2H), 4.11 (bs, 1H ), 5.38-5.40 (m, 1H), 7.11 (s, 1H),7.16-7.36 (m, 15H), 9.45 (s, 1H).

Preparation of Intermediate Ethyl-3-[BOC-L-(Tr-Gln)]-E-Propenoate

Sodium bis(trimethylsilyl)amide (3.38 mL of a 1.0 M solution in THF, 3.3mmol) was added to a solution of triethyl phosphonoacetate (0.732 mL,3.39 mmol) in THF (100 mL) at −78° C., and the resulting solution wasstirred for 20 minutes at that temperature. BOC-L-(Tr-Glutaminal) (1.60g, 3.39 mmol) in THF (20 mL) was added via cannula, and the reactionmixture was stirred for 4 hours at −78° C. then was partitioned between1.0 M HCl (150 mL) and a 1:1 mixture of EtOAc and hexanes (2×150 mL).The organic layers were dried over Na₂SO₄ and concentrated. Purificationof the residue by flash column chromatography (40% EtOAc in hexanes)provided ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (1.53 g, 83%) as a whitefoam: R_(f)=0.60 (50% EtOAc in hexanes); IR (cm⁻¹) 3321, 1710; ¹H NMR(CDCl₃) δ1.27 (t, 3 H, d=7.2 Hz), 1.42 (s, 9H), 1.70-1.78 (m, 1H),1.80-1.96 (m, 1H), 2.35 (t, 2H, J=7.0 Hz), 4.18 (q, 2H, J=7.2 Hz), 4.29(bs, 1H), 4.82-4.84 (m, 1H); 5.88 (dd, 1H, J=15.7, 1.6 Hz), 6.79 (dd,1H, J=15.7, 5.3 Hz), 6.92 (s, 1H), 7.19-7.34 (m, 15H); Anal.(C₃₃H₂₈N₂O₅) C, H, N.

Preparation of ProductEthyl-3-[CBZ-L-Leu-L-Phe-L-(Tri-Gln)]-E-Propenoate

Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.224 g, 0.422 mmol) wasdissolved in 1,4-dioxane (3 mL) and cooled to 0° C. A solution of HCl in1,4-dioxane (4.0 M, 3 mL, 12 mmol) was added dropwise, and the reactionsolution was allowed to arm to room temperature. After being stirred for2 hours, the solution was diluted with 1:1 CH₂Cl₂/EtOAc (50 mL) andadded to a solution of NaOH (16 mmol) in saturated aqueous NaHCO₃ (50mL). After vigorous shaking, the phases were separated, and the aqueousphase was washed 2 more times with 1:1 CH₂Cl₂/EtOAc (50 mL). Thecombined organic phases were dried over Na₂SO₄ and concentrated to give0.164 g (88%) of the crude free amine, which was used without furtherpurification.

The crude amine (0.371 mmol, 1.0 equiv) was dissolved in dry CH₂Cl₂ (5mL). CBZ-L-Leu-L-Phe (0.176 g, 0.427 mmol), 1-hydroxybenzotriazolehydrate (0.81 g, 0.599 mmol), 4-methylmorpholine (0.175 mL, 1.59 mmol),and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.114g, 0.595 mmol) were added sequentially. After being stirred for 18 hoursat 23° C., the reaction mixture was poured into water (40 mL) andextracted with 1:1 CH₂Cl₂/EtOAc (3×50 mL). The combined organic layerswere dried over Na₂SO₄ and were concentrated. The residue was purifiedby flash column chromatography (50% EtOAc in hexanes) to give theproduct (0.163 g, 49%) as a white solid: mp=192-194° C.; IR (KBr) 3295,3049, 1696, 1654 cm⁻¹; ¹H NMR (CDCl₃) δ0.84 (d, 3H, J=6.5 Hz), 0.86 (d,3H, J=6.5 Hz), 1.24-1.32 (m, 1H), 1.28 (t, 3H, J=7.2 Hz), 1.43-1.75 (m,3H), 1.91-2.06 (m, 1H), 2.20-2.38 (m, 2H), 2.93-3.02 (m, 1H), 3.07-3.18(m, 1H), 3.95-4.02 (m, 1H), 4.17 (q, 2H, J=7.2 Hz), 1.43-4.55 (m, 2H),4.82-4.95 (m, 2H), 5.69 (d, 1H, J=15.7 Hz), 6.46 (d, 1H, J=7.5 Hz), 6.60(d, 1H, J=8.1 Hz), 6.69 (dd, 1H, J=15.7, 5.1 Hz), 7.09-7.38 (m, 27 H);Anal. (C₅₁H₅₆N₄O₇) C, H, N.

Example 4 Preparation of Compound 3:Ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofProduct—Ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Compound 2, ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.15 g,0.18 mmol), prepared as described in Example 3, was dissolved in 1:1CH₂Cl₂/TFA (5 mL) at 23° C. and the bright yellow solution was stirred30 minutes, whereupon the solvent was evaporated. CCl₄ (10 mL) wasadded, and the resulting solution was concentrated twice. Addition ofEt₂O (10 mL) to oily residue quickly gave a white precipitate. Afterstirring 10 minutes, the solid was collected by filtration and washedsequentially with acetone (2×10 mL) and Et₂O (2×10 mL) then was dried invacuo to give ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate (0.057 mg,53%) as a white solid: mp=219-221° C.; IR (KBr) 3300, 3065, 1672 cm⁻¹;¹H NMR (DMSO-d₆) δ 0.78 (d, 3H, J=6.8 Hz), 0.82 (d, 3H, J=6.5 Hz), 1.21(t, 3H, J=7.0 Hz), 1.25-1.37 (m, 2H), 1.42-1.54 (m, 1H), 1.58-1.80 (m,2H), 2.02-2.09 (m, 2H), 2.84 (dd, 1H, J=13.2, 8.9 Hz), 2.97 (dd, 1H,J=13.2, 5.8 Hz), 3.93-4.01 (m, 1H), 4.11 (q, 2H, J=7.0 Hz), 4.33-4.52(m, 2H), 4.97 (d, 1H, J=12.3 Hz), 5.04 (d, 1H J=12.3 Hz), 5.64 (d, 1H,J=15.9 Hz), 6.69 (dd, 1H, J=15.9, 5.4 Hz), 6.76 (s, 1H), 7.13-7.37 (m,11H), 7.43 (d, 1H, J=7.8 Hz), 7.99 (d, 1H, J=8.1 Hz), 8.04 (d, 1H, J=8.1Hz); Anal. (C₃₂H₄₂N₄O₇) C, H, N.

Example 5 Preparation of Compound 7:Methyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-Z-Propenoate Preparation ofIntermediate Methyl-3-[BOC-L-(Tr-Gln)]-Z-Propenoate

18-crown-6 (0.867 g, 3.28 mmol) was evaporated from toluene (40 mL) andthen dissolved in dry THF (14 mL) under argon.Bis(2,2,2-trifluoroethyl)(methoxycarbonylmethyl)phosphonate (0.111 mL,0.525 mmol) was added, and the reaction mixture was cooled to −78° C.After dropwise addition of a solution of potassiumbis(trimethylsilyl)-amide in toluene (0.5 M, 1.26 mL, 0.63 mmol), thereaction mixture was stirred for 25 minutes. A solution ofBOC-L-(Tr-glutaminal) (0.310 g, 0.656 mmol) in dry THF (4 mL) was addeddropwise, and, after stirring 1 hour more, saturated aqueous NH₄Cl (2mL) was added. The reaction mixture was allowed to warm to roomtemperature, and the THF was evaporated. Water (10 mL) was added to theresidue, which was then extracted with CH₂Cl₂ (3×30 mL). The combinedorganic phase were dried over Na₂SO₄ and concentrated. The residue waspurified by flash column chromatography (35% EtOAc/hexanes) to give theproduce (0.181 g, 52% ) as a glass: IR (thin film) 3326, 1713, 1690,1666, 1514 cm⁻¹; ¹H NMR (CDCl₃) δ 1.41 (s, 9H), 1.84-1.93 (m, 2H),2.37-2.44 (m, 2H), 3.68 (s, 3H), 5.10 (m, 2H), 5.80 (d, 1H, J=11.8 Hz),6.03 (m, 1H), 6.88 (bs, 1H), 7.18-7.32 (m, 15H).

Preparation of IntermediateMethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-Z-Propenoate

Methyl-3-[BOC-L-(Tr-Gln)]-Z-propenoate (0.143 g, 0.271 mmol) wasdissolved in 1,4-dioxane (3 mL) at room temperature. A solution of HClin 1,4-dioxane (4.0 M, 3 mL) was added dropwise, and the reactionsolution was stirred for 2 hours under an argon balloon. Then thesolvent was evaporated to give the crude amine salt as a glassy residue,which was used without further purification. This amine salt,CBZ-L-Leu-L-Phe (0.112 g, 0.272 mmol), and 1-hydroxybenzotriazolehydrate (0.055 g, 0.40 mmol) were dissolved in dry CH₂Cl₂ (5 mL) underargon at room temperature. 4-Methylmorpholine (0.149 mL, 1.36 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.078 g,0.40 mmol) were then added sequentially. After stirring for 3 hours,water (10 mL) was added, and the mixture was extracted with CH₂Cl₂ (3×30mL). The combined organic phases were dried over Na₂SO₄ andconcentrated. The residue was purified by flash column chromatography(33% acetone in hexanes) to give the product (0.132 g, 59%) as a whitefoam: IR (thin film) 3296, 1708, 1650, 1517 cm⁻¹; Anal. (C₅₀H₅₄N₄O₇) C,H, N.

Preparation of Product—Methyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-Z-Propenoate

Methyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-Z-propenoate (0.110 g, 0.134 mmol)was dissolved in 1:1 CH₂Cl₂/TFA (4 mL), giving a bright yelloe solution,which was stirred for 30 minutes under an argon balloon. CCl₄ (7 mL) wasadded, and the solution was concentrated twice. The residue wastriturated with Et₂O (3 mL) to give a white solid, which was collectedby filtration and dried in vacuo (0.040 g, 51%): mp=185-188° C.; IR(KBr) 3401, 3283, 1719, 1690, 1643, 1538 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.78(d, 3H, J=6.6 Hz), 0.82 (d, 3H, J=6.5 Hz), 1.22-1.38 (m, 2H), 1.43-1.54(m, 1H), 1.58-1.75 (m, 2H), 1.92-2.09 (m, 2H), 2.77-2.90 (m, 2H), 3.65(s, 3H), 3.91-4.00 (m, 1H), 4.37-4.46 (m, 1H), 4.99 (d, 1H, J=12.6 Hz),5.04 (d, 1H, J=12.6 Hz), 5.18-5.25 (m, 1H), 5.79 (d, 1H, J=11.5 Hz),5.92 (dd, 1H, J=11.5, 8.7 Hz), 6.72 (s, 1H), 7.14-7.36 (m, 11H), 7.43(d, 1H, J=8.0 Hz), 7.76 (d, 1H, J=8.1 Hz); Anal. (C₃₁H₄₀N₄O₇) C, H, N.

Example 6 Preparation of Compound 11:Methyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofIntermediate Methyl-3-[BOC-L-(Tr-Gln)]-E-Propenoate

Sodium bis(trimethylsilyl)amide (0.978 mL of a 1.0 M solution in THF,0.978 mmol) was added to a solution of trimethyl phosphonoacetate (0.144mL, 0.890 mmol) in THF (20 mL) at −78° C., and the resulting solutionwas stirred for 15 minutes at that temperature. BOC-L-(Tr-Glutaminal)(0.420 g, 0.889 mmol) in THF (10 mL) was added via cannula, and thereaction mixture was stirred for 2 hours at −78° C., then waspartitioned between 0.5 M HCl (100 mL) and a 1:1 mixture of EtOAc andhexanes (2×100 mL). The organic layers were dried over Na₂SO₄ and wereconcentrated. Purification of the residue by flash column chromatography(gradient elution, 30-40% EtOAc in hexanes) providedmethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.460 g, 96%) as a white solid:mp 110-112° C.; IR (thin film) 3318, 1708, 1665 cm⁻¹; ¹H NMR (CDCl₃) δ1.42 (s, 9H), 1.72-1.82 (m, 1H), 1.91-1.98 (m, 1H), 2.34-2.41 (m, 2H),3.72 (s, 3H), 4.29 (s, br, 1H), 4.78-4.81 (m, 1H), 5.89 (dd, 1H, J=15.6,1.6 Hz), 6.80 (dd, 1H, J=15.6, 5.3 Hz), 6.87 (s, 1H), 7.19-7.33 (m,15H).

Preparation of IntermediateMethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 28 for the preparation ofethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,methyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.157 g, 0.297 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.123 g, 0.298 mmol) toprovide methyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.176 g,72%) as a white foam: ¹H NMR (CDCl₃) δ 0.84 (d, 3H, J=6.7 Hz), 0.86 (d,3H, J=6.7 Hz), 1.45-1.61 (m, 3H), 1.67-1.75 (m, 1H), 1.94-1.96 (m, 1H),2.20-2.35 (m, 2H), 2.95-3.15 (m, 2H), 3.72 (s, 3H, 3.94-4.01 (m, 1H),4.46-4.49 (m, 1H), 4.83-4.93 (m, 3H), 5.72 (d, 1H, J=15.8 Hz), 6.45 (d,1H, J=7.2 Hz), 6.63 (d, 1H, J=8.1 Hz), 6.71 (dd, 1H, J=15.8, 5.1 Hz),7.01-7.38 (m, 27H).

Preparation of Product—Methyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, methyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.087 g,0.106 mmol) was deprotected to providemethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate (0.015 g, 25%) as a whitesolid: mp=220° C. (dec); ¹H NMR (DMSO-d₆) δ 0.79 (d, 3H, J=10.9 Hz),0.81 (d, 3H, J=10.9 Hz), 1.26-1.34 (m, 2H), 1.47-1.49 (m, 1H), 1.61-1.76(m, 2H), 2.06 (t, 2H, J=7.6 Hz), 2.84 (dd, 1H, J=13.5, 9.0 Hz), 2.97(dd, 1H, J=13.5, 5.6 Hz), 3.65 (s, 3H), 3.93-3.97 (m, 1H), 4.38 (s, br,1H), 4.44-4.49 (m, 1H), 4.97 (d, 1H, J=12.5 Hz), 5.04 (d, 1H, J=12.5Hz), 5.68 (d, 1H, J=15.6 Hz), 6.70 (dd, 1H, J=15.6, 5.5 Hz), 6.76 (s,1H), 7.19 (s, br, 7H), 7.34 (s, br, 4H), 7.44 (d, 1H, J=7.5 Hz), 7.99(d, 1H, J=8.1 Hz), 8.05 (d, 1H, J=8.1 Hz).

Example 7 Preparation of Compound 13:4-(CBZ-L-Leu-L-Phe-L-Gln)-E-3-Butene-2-one Preparation of IntermediateCBZ-L-Leu-L-Phe-L-(Tr-Gln)-N(Me)OMe

BOC-L-(Tr-Gln)-N(Me)OMe (0.807 g, 1.52 mmol) was dissolved in1,4-dioxane (4.5 mL) at room temperature. A solution of HCl in1,4-dioxane (4.0 M, 4.5 mL) was added dropwise, and the reactionsolution was stirred for 2.5 hours under an argon balloon. The solventwas evaporated to give the crude amine salt as a white foam, which wasused without further purification. This amine salt, CBZ-L-Leu-L-Phe(0.626 g, 1.52 mmol) and 1-hydroxybenzotriazole hydrate (0.308 g, 2.28mmol) were stirred in dry CH₂Cl₂ (12 mL) under argon at roomtemperature. 4-Methylmorpholine (0.840 mL, 7.64 mmol), and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.436 g,2.27 mmol) were added squentially. After stirring for 3 hours, thereaction solution was poured into water (25 mL), and the aqueous layerwas extracted 3 times with CH₂Cl₂ (70 mL, 40 mL, and 30 mL). Thecombined organic phases were dried over Na₂SO₄ and concentrated. Theresidue was purified by flash column chromatography (40% acetone inhexanes) to give the product (0.826 g, 66%) as a white foam: IR (thinfilm) 3300, 1643, 1525 cm⁻¹.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(Tr-Glutaminal)

CBZ-L-Leu-L-Phe-L-(Tr-Gln)-N(Me)OMe (0.768 g, 0.930 mmol) was dissolvedin dry THF (12 mL) under argon and cooled to −78° C. A solution ofdiisobutylaluminum hydride in toluene (1.5 M, 2.17 mL, 3.26 mmol) wasadded dropwise. After stirring 3 hours, methanol (0.7 mL) was addedslowly, followed by 1 N HCL (1 mL). The reaction mixture was allowed towarm to nearly room temperature and was then diluted with 5:1CH₂Cl₂/EtOAc (120 mL). The resulting mixture was washed with 1 N HCl(2×15 mL), half-saturated NaHCO₃ (15 mL) and brine (25 mL). The organicphase was dried over MgSO₄ and concentrated to give the product as anoff-white foam (0.606 g, 85%), which was used without furtherpurification. An analytical sample was purified by column chromatography(36% acetone in hexanes): IR (thin film) 3295, 1708, 1660, 1531 cm⁻¹; ¹HNMR (CDCl₃) δ 0.80 (d, 3H, J=6.2 Hz), 0.87 (d, 3H, J=6.4 Hz), 1.27-1.59(m, 3H), 1.71-1.83 (m, 1H), 2.07-2.15 (m, 1H), 2.22-2.29 (m, 2H), 2.96(dd, 1H, J=13.7, 7.4 Hz), 3.08 (dd, 1H, J=13.7, 6.2 Hz), 3.99-4.08 (m,1H), 4.11-4.20 (m, 1H), 4.55-4.64 (m, 1H), 4.92 (bs, 2H), 5.17 (d, 1H,J=6.7 Hz), 6.70 (d, 1H, J=7.4 Hz), 7.08-7.35 (m, 27H), 9.26 (s, 1H);Anal. (C₄₇H₅₀N₄O₆) C, H, N.

Preparation of Intermediate4-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Butene-2-one

CBZ-L-Leu-L-Phe-L-(Tr-Glutaminal) (0.605 g, 0.789 mmol) and1-triphenylphosphoranylidene-2-propanone (0.251 g, 0.788 mmol) werestirred in dry THF (7 mL) at room temperature, under argon, giving ayellow solution. After stirring 20 hours, the solvent was evaporated,and the residue was purified by flash column chromatography (36% acetonein hexanes) to give the product (0.425 g, 67%) as a white foam: IR (thinfilm) 3299, 1666, 1519 cm⁻¹.

Preparation of Produce-4-(CBZ-L-Leu-L-Phe-L-Gln)-E-3-Butene-2-one

This compound was prepared in 54% yield from4-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-3-butene-2-one using the proceduredescribed in Example 26 for the prepartion of compound 14,3-(CBZ-L-Leu-L-Phe-DL-Gln)-E-propenonitrile: mp=194-196° C. (dec); IR(KBr) 3413, 3284, 1684, 1643, 1537 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.79 (d, 3H,J=6.6 Hz), 0.82 (d, 3H, J=6.6 Hz), 1.23-1.39 (m, 2H), 1.44-1.55 (m, 1H),1.60-1.84 (m, 2H), 2.05-2.12 (m, 2H), 2.17 (s, 3H), 2.84 (dd, 1H,J=13.6, 8.7 Hz), 2.99 (dd, 1H, J=13.6, 5.7 Hz), 3.93-4.02 (m, 1H,4.34-4.44 (m, 1H, 4.46-4.55 (m, 1H), 4.98 (d, 1H, J=12.6 Hz), 5.04 (d,1H, J=12.6 Hz), 5.84 (d, 1H, J=16.0 Hz), 6.64 (dd, 1H, J=16.0, 5.4 Hz),6.77 (s, 1H), 7.15-7.37 (m, 11H), 7.43 (d, 1H, J=7.9 Hz), 7.99 (d, 1H,J=8.1 Hz), 8.06 (d, 1H, J=8.1 Hz); Anal. (C₃₁H₄₀N₄O₆) C, H, N.

Example 8 Preparation of Compound 5:Ethyl-3-[CBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Ala]-E-PropenoatePreparation of Intermediate CBZ-L-[N-(4-Chlorobutyryl)-amino]-Ala-OMe

Acetyl chloride (19.6 g, 250 mmol) was slowly added to a solution ofMeOH (300 mL) at 0° C. After 10 minutes, CBZ-L-amino-Ala (10 g., 42mmol) was added, and the reaction was allowed to stir for 12 hours atroom temperature. Removal of solvent under vacuum provided 13.5 g ofcrude CBZ-L-amino-Ala-OMe as the hydrochloride salt. The crude ester wastaken up in 200 mL CH₂Cl₂, to which was added Et₃N (10.6 g, 105 mmol)and then 4-chlorobutyryl chloride (7.1 g, 50.4 mmol) at 0° C. Thereaction was allowed to warm to room temperature and was stirred for 4hours. At this time the reaction mixture was added to brine. The organiclayer was extracted, washed with 1 N HCl, brine, dried over MgSO₄, andconcentrated yielding 19 g of crude material. The material was purifiedby flash column chromatography (50% EtOAc-hexanes), giving an 87% yieldof product. ¹H NMR (CDCl₃) δ 2.07 (m, 2H), 2.35 (t, 2H, J=7.0 Hz), 3.57(t, 2H, J=6.3 Hz), 3.67 (t, 2H, J=5.9 Hz), 3.77 (s, 3H), 4.45 (m, 1H),5.12 (s, 2H), 5.84 (d, 1H, J=6.3 Hz), 6.00 (bs, 1H), 7.37 (s, 5H).

Preparation of Intermediate CBZ-L-[N-(2-pyrrolidinone)]-Ala-OMe

A solution of CBZ-L-[N-(4-chlorobutyryl)-amino]-Ala-OMe (14.6 g, 39mmol) in DMF (400 mL) was cooled to 0° C. To the solution was added NaH(1.87 g of a 60% dispersion in oil, 46.8 mmol), and the mixture wasstirred at room temperature for 4 hours. The DMF was removed under highvacuum, and the residue was taken up in EtOAc, washed with 1 N HCl,saturated aqueous NaHCO₃, brine, dried over MgSO₄ and concentrated. Thematerial was purified by flash column chromatography (100% EtOAc),giving 7.0 g (56%) of product. ¹H NMR δ (CDCl₃) 1.97 (m, 2H), 2.35 (m,2H), 3.36 (m, 1H), 3.40-3.60 (m, 3H), 3.77 (s, 3H), 4.52 (m, 1H), 5.13(d, 2H, J=5.6 Hz), 5.83 (d, 1H, J=6.3 Hz), 7.37 (m, 5H).

Preparation of Intermediate L-[N-(2-pyrrolidinone)]-Ala-OMeHCl

This compound was prepared from CBZ-L-[N-(2-pyrrolidinone)]-Ala-OMe bycatalytic hydrogenation as described in Example 2 for the preparation ofL-(N-Ac-amino)-alaninol, except methanolic HCl was used in order toisolate the product as the HCl salt. ¹H NMR (CDCl₃), δ 2.03 (m, 2H),2.39 (m, 2H), 3.14 (bs, 2H), 3.40-3.70 (m, 5H), 3.75 (s, 3H).

Preparation of IntermediateCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Ala-OMe

This compound was prepared from CBZ-L-Leu-L-Phe andL-[N-(2-pyrrolidinone)]-Ala-OMe HCl using the procedure described inExample 1 for the preparation of CBZ-L-Leu-L-Phe-L-methioninol. ¹H NMR(CDCl₃), δ 0.89 (m, 6H), 1.36 (m, 2H), 1.56 (m, 1H), 1.61 (m, 2H), 2.04(m, 3H), 2.31 (m, 2H), 3.07-3.70 (m, 6H), 3.75 (s, 3H), 4.11 (m, 1H),4.71 (m, 1H), 5.13 (bs, 1H), 5.18 (bs, 1H), 6.76-6.88 (m, rotomers, 1H),7.10-7.45 (m, 10H).

Preparation of IntermediateCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Alaninol

This compound was prepared by the reduction ofCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Ala-OMe with NaBH₄ and LiClusing the procedure described in Example 2 for the preparation ofCBZ-L-(N-Ac-amino)-alaninol.

Preparation of IntermediateCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Alaninal

This compound was prepared fromCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-alaninol using the proceduredescribed in Example 1 for the preparation ofCBZ-L-Leu-L-Phe-L-methioninal (sulfoxide). Anal. (C₃₀H₃₈N₄O₆·1.4 H₂O) C,H, N.

Preparation ofProduct—Ethyl-3-[CBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-Ala]-E-Propenoate

This compound was prepared fromCBZ-L-Leu-L-Phe-L-[N-(2-pyrrolidinone)]-alaninal and(carbethoxymethylene)triphenylphosphorane using the procedure describedin Example 1 for the preparation of compound 12,ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate. ¹H NMR (DMSO-d₆)δ 0.80 (d, 6H, J=7.0 Hz), 0.95-1.40 (m, 7H), 1.49 (m, 1H), 1.82 (m, 2H),2.12 (m, 2H), 2.60-3.10 (m, 2H), 3.20 (m, 2H), 3.81 (m, 1H), 4.00 (m,1H), 4.10 (m, 2H), 4.49 (m, 1H), 4.72 (m, 1H), 5.01 (bs, 1H), 5.70 (d,0.5H-rotomer-H=16.5 Hz), 5.97 (d, 0.5H-rotomer-J=16.5 Hz), 6.70 (d,0.5H-rotomer-J=16.5 Hz), 6.80 (d, 0.5H-rotomer-J=16.5 Hz), 7.20 (d, 2H,J=7.4 Hz), 7.34 (m, 3H, 7.60 (m, 5H), 8.04 (m, 1H, 8.23 (m, 1H), HRMScalcd for C₃₄H₄₄N₄O₇+Cs 753.2264 (M+Cs), found 753.2295.

Example 9 Preparation of Compound 16:Ethyl-3-[CBZ-L-Leu-L-Phe-L-(N-carbamyl-amino)-Ala]-E-PropenoatePreparation of Intermediate CBZ-L-(N-BOC-amino)-Ala

To a stirred solution of NaOH (1.23 g, 30.76 mmol) in 36 mL of H₂O and24 mL tert-butanol was added CBZ-L-amino-Ala (7.15 g, 30 mmol). To thissolution was added di-tert-butyl dicarbonate (6.88 g, 31.5 mmol).Stirring was continued at room temperature for 12 hours, at which timethe solution was washed with pet. ether (2×150 mL). The organic layerswere washed with saturated aqueous NaHCO₃ (3×20 mL), and the aqueouslayers were combined and acidified at 0° C. with 25% aqueous KHSO₄ to pH2-3. This milky white mixture was then extracted with a large excess ofEt₂O, dried over anhydrous Na₂SO₄, and concentrated to yield 9.13 g(90%) of product as a white solid, which was used without furtherpurification. ¹H NMR (DMSO-d₆) δ 1.35 (s, 9H), 3.21 (m, 2H), 4.05 (m,1H), 5.02 (s, 2H), 6.83 (bt, 1H, J=6.6 Hz), 7.34 (m, 5H), 7.41 (d, 1H,J=8.1 Hz), 12.65 (bs, 1H). This compound was further characterized asits corresponding methyl ester.

Preparation of Intermediate CBZ-L-(N-BOC-amino)-Ala-OMe

A solution of diazomethane in Et₂O, generated fromN-methyl-N-nitroso-p-toluenesulfonamide (7.7 g, 36.0 mmol), 70 mL Et₂O,16 mL EtOH, 12 mL H₂O and KOH (7.65 g, 13.6 mmol) was carefullydistilled into a stirred solution of CBZ-L-(N-BOC-amino)-Ala (7.8 g,23.0 mmol) in 50 mL Et₂O and 10 mL EtOH at 0° C. The yellow solution wasstirred for 30 minutes. The cold solution was then brought to roomtemperature, and argon was bubbled into the reaction flask to remove anyexcess diazomethane. After the solution turned colorless, it wasconcentrated to give the methyl ester as a white solid in quantitativeyield. mp=72-74° C.; IR (KBr) 3418, 3331, 3005, 2955, 1753, 1724, 1676,1552, 1525, 1298, 1045, 699 cm⁻¹; ¹H NMR (CDCl₃) δ 1.41 (s, 9H), 3.55(m, 2H), 3.76 (s, 3H), 4.40 (m, 2H), 4.82 (m, 1H), 5.11 (s, 2H), 5.77(m, 1H), 7.35 (m, 5H). Anal. (C₁₇H₂₄N₂O₆) C, H, N.

Preparation of Intermediate CBZ-L-(N-BOC-amino)-Alaninol

Using the borohydride reduction procedure described in Example 2 for theprepartion of CBZ-L-(N-Ac-amino)-alaninol, CBZ-L-(N-BOC-amino)-Ala-OMewas converted to the corresponding alcohol and isolated in 96% yieldwithout column chromatography purification. mp=116-119° C.; IR (KBr)3327, 3277, 3065, 2976, 1699, 1682, 1543, 1315, 1250, 1062, 1001, 696cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.35 (s, 9H), 2.90-3.10 (m, 4H), 3.55 (m 1H),4.60 (bt, 1H, J=5.5 Hz), 4.99 (s, 2H), 6.72 (bt, 1H, J=5.5 Hz), 6.86 (d,1H, J=8.1 Hz), 7.34 (m, 5H). Anal. (C₁₆H₂₄N₂O₆) C, H, N.

Preparation of Intermediate L-(N-BOC-amino)-Alaninol

Using the hydrogenation procedure described in Example 2 for thepreparation of L-(N-Ac-amino)-alaninol, the CBZ group was removed fromCBZ-L-(N-BOC-amino)-alaninol to give the amino alcohol in 98% yield.mp=61-64° C.; IR (KBr) 3362, 2980, 2935, 1680, 1534, 1370, 1287, 1175,1059, 642 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.36 (s, 9H), 2.64 (m, 1H), 2.72 (m,1H), 2.93 (m, 1H), 3.13 (m, 1H), 3.32 (m, 2H), 4.45 (bs, 1H), 6.67 (bs,1H); Anal. (C₂H₁₂N₂O₃) C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(N-BOC-amino)-Alaninol

This compound was prepared from L-(N-BOC-amino)-alaninol andCBZ-L-Leu-L-Phe using the coupling procedure described in Example 2 forthe preparation of CBZ-L-Leu-L-Phe-L-(N-Ac-amino)-alaninol. The reactionmixture was purified by flash column chromatography (5% saturatedanhydrous NH₃ in MeOH/CH₂Cl₂) to give a white solid in 90% yield. IR(KBr) 3420, 3327, 3289, 3032, 2953, 1694, 1643, 1535, 1284, 1036, 696cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.80 (dd, 6H, J=11.2, 6.4 Hz), 1.35 (s, 9H),1.55 (m 2H), 1.72 (m, 1H), 2.89 (m, 2H, 3.19 (m, 2H), 3.78 (m, 1H), 3.92(m, 1H), 4.44 (m, 1H), 4.62 (t, 1H, J=5.5 Hz), 5.01 (d, 2H, J=5.9 Hz),6.63 (bt, 1H, J=5.5 Hz), 7.18 (m, 5H), 7.34 (m, 5H), 7.45 (d, 1H, J=8.1Hz), 7.60 (d, 1H, J=7.7 Hz), 7.85 (d, 1H, J=8.1 Hz). Anal. (C₃₁H₄₄N₄O₇)C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(N-BOC-amino)-Alaninol

This compound was prepared in 90% yield as white solid fromCBZ-L-Leu-L-Phe-L-(N-BOC-amino)-alaninol using the procedure describedin Example 1 for the preparation of CBZ-L-Leu-L-Phe-L-methioninal(sulfoxide). The product was used immediately without furtherpurification. The product existed as a mixture of aldehyde and aldehydehydrate. IR (KBr) 3299, 3067, 2959, 2934, 1696, 1647, 1535, 1254, 1171,747, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.80 (dd, 6H, J=9.0, 6.8 Hz), 1.35 (s,9H), 1.41 (m, 2H), 1.69 (m, 1H), 2.80-3.03 (m, 2H), 3.29 (m, 2H), 3.97(m, 1H), 4.10 (m, 1H), 4.60 (m, 1H), 5.00 (s, 2H), 5.56 (d, J=7.4 Hz,hydrate), 6.78 (t, 1H, J=6.3 Hz), 7.20 (m, 5H), 7.33 (m, 5H), 7.40 (d,1H, J=8.1 Hz), 7.97 (d, 1H, J=8.1 Hz), 8.39 (d, 1H, J=6.6 Hz), 9.26 (s,1H); HRMS calcd for C₃₁H₄₂N₄O₇+Cs 715.2108 (M+Cs), found 715.2133. Anal.(C₃₁H₄₂N₄O₇0.5 H₂O) C, H, N.

Preparation of IntermediateEthyl-3-[CBZ-L-Leu-L-Phe-L-(N-BOC-amino)-Ala]-E-Propenoate

This compound was prepared in approximately 40% yield as a white foamingsolid from CBZ-L-Leu-L-Phe-L-(N-BOC-amino)-alaninol and(carbethoxymethylene)-triphenyl-phosphorane using the proceduredescribed in Example 1 for the preparation of compound 12,ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate. The product waspartially purified (impure with triphenylphospine oxide as determined byNMR) by flash column chromatography (4% MeOH/CH₂Cl₁). ¹H NMR (DMSO-d₆) δ0.80 (dd, 6H, J=9.6, 6.3 Hz), 1.19 (t, 3H, J=6.8 Hz), 1.34 (s, 9H),1.45-1.70 (m, 3H), 2.82-3.05 (m, 4H), 3.99 (m, 1H), 4.08 (q, 2H, J=7.0Hz), 4.46 (m, 2H), 5.01 (m, 2H), 5.64 (d, 1H, J=16.2 Hz), 6.61 (dd, 1H,J=16.2, 5.5 Hz), 6.85 (bt, 1H, J=5.2 Hz), 7.18 (m, 5H), 7.34 (m, 5H),7.42 (d, 1H, J=5.5 Hz), 7.96 (d, 1H, J=7.4 Hz), 8.01 (d, 1H, J=7.4 Hz);HRMS calcd for C₃₅H₁₈N₄O₆+Na 675.3370 (M+Na), found 675.3363.

Preparation of IntermediateEthyl-3-(CBZ-L-Leu-L-Phe-L-amino-Ala)-E-Propenoate

To a stirred solution ofethyl-3-[CBZ-L-Leu-L-Phe-L-(N-BOC-amino)-Ala]-E-propenoate (0.14 g,0.215 mmol) in 12 mL CH₂Cl₂, cooled to 0° C., was added 0.65 mL TFAdropwise. The reaction was followed by TLC (silica, 10% MeOH/CH₂Cl₂)until there was a disappearance of starting material. At this time thereaction mixture was taken up in 100 mL EtOAc and washed with saturatedNaHCO₃ (3×10 mL). The organic layer was then washed with H₂O thensaturated brine and dried over anhydrous Na₂SO₄. Concentration of thesolution gave a residue, which was purified by flash columnchromatography (8% MeOH/CH₂Cl₂) to give a beige foam in 84% yield. ¹HNMR (DMSO-d₆) δ0.80 (dd, 6H, J=9.4, 6.8 Hz), 1.22 (t, 3H, J=7.2 Hz),1.31 (m, 2H), 1.51 (m, 1H), 2.64 (m, 2H), 2.91 (m, 2H), 3.99 (m, 1H),4.10 (q, 2H, J=7.4 Hz), 4.36 (m, 1H), 4.49 (m, 1H), 5.02 (m, 2H), 5.60(d, 1H, J=16.2 Hz), 6.76 (dd, 1H, J=15.6, 5.0 Hz), 7.20 (m, 5H), 7.34(m, 5H), 7.46 (d, 1H, J=7.0 Hz), 7.95 (d, 1H, J=8.5 Hz), 8.05 (d, 1H,H=5.9 Hz); MS calcd for C₃₅H₄₃N₄O₆+H 553 (M+H), found 553.

Preparation ofProduct—Ethyl-3-[CBZ-L-Leu-L-Phe-L-(N-carbamyl-amino)-Ala]-E-Propenoate

To a stirred solution of bis (4-nitrophenyl) carbonate (66 mg, 0.22mmol) in 2 mL CH₂Cl₂, was added a solution ofethyl-3-[CBZ-L-Leu-L-Phe-L-amino-Ala]-E-propenoate (0.10 g, 0.18 mmol)in 2 mL CH₂Cl₂. The mixture was stirred for 3 hours at which time 2 mLof saturated anhydrous methanolic ammonia was added. The yellow solutionwas allowed to stir for 30 minutes longer, diluted with 100 mL CH₂Cl₂,and washed repeatedly with 1N NaOH to remove 4-nitrophenol. The organiclayer was washed with dilute HCl, H₂O, and brine, and dried overanhydrous Na₂SO₄. This solution was concentrated, and the residue wassubjected to flash column chromatography (5% MeOH/CH₂Cl₂) to yield awhite solid in 20% yield. IR (KBr) 3470, 3291, 2978, 2926, 1715, 1645,1539, 1281, 1045, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.81 (dd, 6H, J=9.0, 6.8Hz), 1.21 (t, 3H, J=7.0 Hz), 1.30 (m, 2H), 1.48 (m, 1H), 2.92 (m, 2H),3.10 (m, 2H), 3.97 (m, 1H), 4.10 (q, 2H, J=7.0 Hz), 4.40 (m, 2H), 5.01(m, 2H), 5.54 (bs, 2H), 5.61 (d, J=16.5 Hz), 6.04 (t, 1H, J=7.7 Hz),6.71 (dd, J=15.8, 5.2 Hz), 7.20 (m, 5H), 7.34 (m, 5H), 7.46 (d, 1H,J=7.4 Hz), 8.01 (d, 1H, J=7.0 Hz), 8.11 (d, 1H, J=8.5 Hz); HRMS calcdfor C₃₁H₄₁N₅O₇+Cs 728.2060 (M+Cs), found 728.2078 Anal. (C₃₁H₄₁N₅O₇) C,H, N.

Example 10 Preparation of Compound 17:Isopropyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofIntermediate 3-[BOC-L-(Tr-Gln)]-E-Propenoic Acid

Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (1.874 g, 3.46 mmol), prepared asdescribed in Example 3, was taken up in 20 mL EtOH and treated with 1NNaOH solution (7.95 mL, 7.95 mmol) dropwise, via addition funnel, over 2hours. The resulting solution was stirred at room temperature for 1.5hours, whereupon the reaction mixture was poured into water and washedwith ether. The aqueous layer was acidified to pH 3 with 1N HCl, andextracted 3 times with EtOAc. The oragnic phase was separated and driedover MgSO₄ and concentrated to provide 3-[BOC-L-(Tr-Gln)]-E-propenoicacid (1.373 g, 77%) as an off-white foam. No further purification wasneeded: IR (thin film) 3315, 1698, 1666 cm⁻¹; ¹H NMR (CDCl₃) δ 1.42 (s,9H), 1.76 (m, 1H), 1.83-1.98 (m, 1H), 2.37 (t, 2H, J=7.0 Hz), 4.30 (m,1H), 4.88 (m, 1H), 5.85 (d, 1H, J=15.3 Hz), 6.86 (dd, 1H, J=15.5, 5.1Hz), 6.92 (s, 1H), 7.25 (m, 15H).

Preparation of Intermediate Isopropyl-3-[BOC-L-(Tr-Gln)-E-Propenoate

3-[BOC-L-(Tr-Gln)]-E-Propenoic acid (0.500 g, 0.973 mmol), isopropanol(0.008 mL, 1.07 mmol), and 4-dimethylaminopyridine (0.012 g, 0.0973mmol) were taken up in 10 mL dry CH₂Cl₂ and treated with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.196 g,1.07 mmol), The resulting solution was stirred at room temperatureovernight, concentrated in vacuo, and purified by flash column with 50%EtOAc/hexanes to provide isopropyl-3-[BOC-L-(Tr-Gln)]-E-propenoate(0.106 g, 20%) as a white foam: R=0.8 (50% EtOAc/hexanes); IR 3320, 1711cm⁻¹; ¹H NMR (CDCl₃) δ 1.25 (d, 6H, J=6.23 Hz), 1.43 (s, 9H), 1.72 (m,1H), 1.96 (m, 1H), 2.37 (t, 2H, J=7.16 Hz), 4.30 (bs, 1H), 4.74 (m, 1H),5.05 (m, 1H), 5.86 (dd, 1H, J=15.9, 5.0 Hz), 6.78 (dd, 1H, J=15.6, 5.0Hz), 6.89 (bs, 1H), 7.26 (m, 15H); Anal. (C₃₄H₄₀N₂O₅) C, H, N.

Preparation of IntermediateIsopropyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate

Isopropyl-3-]BOC-L-[BOC-L-(Tr-Gln)]-E-propenoate (0.087 g, 0.191 mmol)was deprotected and coupled with CBZ-L-Leu-L-Phe (0.079 g, 0.191 mmol)using the procedure described in Example 3 for the preparation ofethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate, to provide the product (0.064 g,40%) as a white foam: R_(ƒ)=0.7 (50% EtOAc/hexanes); IR (thin film)3283, 1707 cm⁻¹; ¹H NMR (CDCl₃) δ 0.86 (m, 6H), 1.03 (m, 1H), 1.23 (m,6H), 1.72 (m, 1H), 1.96 (m, 1H), 2.28 (m, 2H), 2.54 (m, 1H), 2.70 (m,1H), 2.78 (m, 1H), 2.95-3.25 (m, 4H), 3.99 (m, 1H), 4.85-5.13 (m, 4H),5.66 (d, 1H, J=15.9 Hz), 6.45 (d, 1H, J=7.5 Hz), 6.55 (d, 1H, J=7.5 Hz),6.68 (m, 1H), 7.12-7.36 (m, 25H); MS (M+Cs) 983.

Preparatoin of Product—Isopropyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate,isopropyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.059 g, 0.0694mmol) was deprotected to provide the product (0.024 g, 57%) as a whitesolid: mp=180-182° C.; R_(ƒ)=0.6 (10% MeOH/CHCl₃); IR (KBr) 3272, 1705cm⁻¹; ¹H NMR (DMSO-d_(6) δ) 0.70 (m, 1H), 0.80 (dd, 6H, J=10.6, 6.5 Hz),1.21 (dd, 6H, J=6.2, 2.5 Hz), 1.32 (m, 1H), 1.70 (m, 1H), 2.05 (t, 2H,J=7.6 Hz), 2.83 (m, 1H), 2.97 (m, 1H), 3.99 (m, 1H), 4.37-4.49 (m, 4H),4.91-5.06 (m, 4H), 5.60 (d, 1H, J=15.3 Hz), 6.67 (dd, 1H, J=15.6, 5.6Hz), 6.76 (bs, 1H), 7.19 (m, 5H), 7.34 (m, 5H), 7.44 (d, 1H, J=7.2 Hz),8.01 (m, 2H); Anal. (C₃₃H₄₄N₄O₇·1.0 CH₂Cl₂) C, H, N.

Example 11 Preparation of Compound 18:Cyclopentyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofIntermediate Cyclopentyl-3-[BOC-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 10 for the preparation ofisopropyl-3-[BOC-L-(Tr-Gln)]-E-propenoate,3-[BOC-L-(Tr-Gln)]-E-propenoic acid (0.50 g, 0.973 mmol) was coupledwith cyclopentanol (0.1 mL, 1.07 mmol) to providecyclopentyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.123 g, 22%) as a whitefoam: R_(ƒ)=0.7 (EtOAc/hexanes); IR (thin film) 3319, 1708 cm⁻¹; ¹H NMR(CDCl₃) δ 1.27 (m, 2H), 1.44 (s, 9H), 1.59-1.89 (m, 8H), 2.38 (t, 2H,J=7.2 Hz), 4.32 (bs, 1H), 4.55 (m, 1H), 5.22 (m, 1H), 5.87 (d, 1H,J=15.6 Hz), 6.77 (dd, 1H, J=15.1, 4.1 Hz), 6.90 (bs, 1H), 7.20-7.33 (m,15H).

Preparation of IntermediateCyclopentyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3[CBZ-L-Phe-L-(Tr-Gln)]-E-propenoate,cyclopentyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.077 g, 0.160 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.068 g, 0.160 mmol) toprovide cyclopentyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.052g, 36%) as a white foam: R_(ƒ)=0.4 (50% EtOAc/hexanes); IR (thin film)3401, 3319, 1708 cm⁻¹; ¹H NMR (CDCl₃) δ 0.84 (m, 6H), 1.05 (m, 1H), 1.28(m, 1H), 1.46-1.71 (m, 9H), 1.85 (m, 1H), 2.28 (m, 2H), 2.98-3.12 (m,4H), 3.99 (m 1H), 4.47 (m, 2H), 4.83-5.21 (m, 4H), 5.65 (d 1H, J=15.9Hz), 6.50 (d, 1H, J=7.2 Hz), 6.59 (d, 1H, J=8.1 Hz), 6.65 (dd, 1H,J=15.9, 5.4 Hz), 7.04-7.35 (m, 25H); MS (M+Cs) 1009.

Preparation ofProduct—Cyclopentyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate,cyclopentyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.052 g, 0.059mmol) was deprotected to provide the product (0.014 g, 36%) as a whitesolid: mp=182-185° C.; R_(ƒ)=0.5 (10% MeOH/CHCl₃); IR (thin film) 3389,3295, 1707 cm⁻¹; ¹H NMR (Acetone-d₆) δ 0.85 (dd, 6H, J=10.6, 6.5 Hz),1.08 (m, 1H), 1.48 (m, 1H), 1.60-1.70 (m, 11H), 1.89 (m, 1H), 2.22 (m,2H), 2.96 (m, 1H), 3.18 (dd, 1H, J=13.9, 5.8 Hz), 4.00 (d, 1H, J=6.8Hz), 4.08 (m, 1H), 4.59 (m, 2H), 4.97-5.16 (m, 4H), 5.76 (d, 1H, J=15.3Hz), 6.71 (m, 2H), 7.15-7.41 (m, 10H), 7.51 (d, 1H, J=7.8 Hz); HRMScalcd for C₃₅H₄₆N₇O₇+Cs 767.2421 (M+Cs) found 767.2435.

Example 12 Preparation of Compound 19:Cyclopentylmethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofIntermediate Cyclopentylmethyl-3-[BOC-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 10 for the preparation ofisopropyl-3-[BOC-L-(Tr-Gln)]-E-propenoate,3-[BOC-L-(Tr-Gln)]-E-propenoic acid (0.50 g, 0.973 mmol) was coupledwith cyclopentylmethanol (0.12 mL, 1.07 mmol) to provide this ester(0.298 g, 51%) as a pale yellow oil: R_(ƒ)=0.7 (50% EtOAc/hexanes); IR(thin film) 3336, 1707 cm⁻¹; ¹H NMR (CDCl₇) δ 1.28 (m, 2H), 1.43 (s,9H), 1.54-1.62 (m, 5H), 1.72-1.78 (m, 4H), 2.37 (t, 2H, J=7.2 Hz), 4.01(d, 2H, J=7.2 Hz), 4.31 (bs, 1H), 4.78 (m, 1H), 5.90 (dd, 1H, J=15.9,1.6 Hz), 6.80 (dd, 1H, J=15.9, 5.3 Hz), 6.90 (bs, 1H), 7.19-7.34 (m,15H); Anal (C₃₇H₄₄N₂O₅) C, H, N.

Preparation of IntermediateCyclopentylmethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,cyclopentylmethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.150 g, 0.310 mmol)was deprotected and coupled with CBZ-L-Leu-L-Phe (0.128 g, 0.310 mmol)to provide the product (0.062 g, 22%) as an off-white foam: R_(f)=0.4(50% EtOAc/hexanes); IR (thin film) 3413, 3295, 1708 cm⁻¹, ¹H NMR(CDCL_(t)) δ 0.84 (m, 6H), 1.05 (m, 1H), 1.46-1.65 (m, 10H), 1.74 (m,1H), 2.25 )m, 2H), 2.93-3.11 (m, 4H), 3.93-4.02 (m, 3H), 4.20 (m, 1H),4.48 (m, 1H), 4.86-5.11 (m, 4H), 5.70 (d, 1H, J=15.0 Hz), 6.46 (d, 1H,J=6.9 Hz), 6.54 (d, 1H, J=8.4 Hz), 6.70 (m, 1H), 6.78 (m, 1H), 7.14-7.36(m, 25H); MS (M+Cs) 1023.

Preparation of Product -Cyclopentylmethyl-3-CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate,cyclopentylmethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.062 g,0.070 mmol) was deprotected to provide compound 11 (0.021 g, 47%) as awhite solid: mp=145-148° C.; R_(f)=0.4 (10% MeOH/CHCl₃); IR (thin film)3401, 3295, 1713 cm⁻¹, ¹H NMR (acetone-d₆) δ 0.86 (dd, 6H, J=10.6, 6.5Hz), 1.09 (m, 1H), 1.20-1.85 (m, 13H), 2.21 (m, 2H), 2.99 (m, 1H), 3.18(m, 1H), 3.99 (m, 2H), 4.10 (m, 2H), 4.59 (m, 2H), 4.98-5.16 (m, 4H),5.83 (d, 1H, J=14.6 Hz), 6.67-6.98 (m, 2H), 7.20-7.45 (m, 10H), 7.55 (m,1H); HRMS calcd for C₃₆H₄₈N₇O₇+Cs 781.2577 (M+Cs) found 781.2559.

Example 13 Preparation of Compound 21:1-Pyrrolidin-1-yl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenone Preparation ofIntermediate 1-Pyrrolidin-1-yl-3-[BOC-L-(Tr-Gln)]-E-Propenone

3-[BOC-L-(Tr-Gln)]-E-Propenoic acid (1.09 g, 2.12 mmol) was coupled withpyrrolidine (0.18 mL, 2.12 mmol) by dissolving both in 30 mL dry Ch₂Cl₂and treating with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (0.610 g, 3.18 mmol), 1-hydroxybenzotriazole hydrate(0.430 g, 3.18 mmol), Et₃N (1.18 mL, 8.48 mmol) and stirring at roomtemperature overnight. The reaction mixture was poured into 50 mL 1NHCl, and the layers were separated. The organic layer was washed with 1NHCl and then a saturated NaHCO₃ solution. The organic layer was driedover MgSO₄ and concentrated to give a yellow residue, which was thensubjected to column chromatography using a 5% MeOH/CHCl₃ to yield theproduct (0.661 g, 55%) as a white foam: R_(f)=0.5 (5% MeOH/CHCl₃); IR(thin film) 3291, 1696 cm⁻¹; ¹H NMR (CDCl₃) δ 1.42 (s, 9H), 1.89 (m,6H), 2.37 (m, 2H), 3.44-3.53 (m, 4H), 4.28 (bs, 1H), 4.82 (d, 1H, J=7.8Hz), 6.17 (dd, 1H, J=15.3, 1.6 Hz), 6.71 (dd, 1H, J=15.4, 6.1 Hz), 6.93(bs, 1H), 7.19-7.32 (m, 15H); Anal (C₃₅H₄₁N₃O₂Cl_(z)) C, H, N.

Preparation of Intermediate1-Pyrrolidin-1-yl-3-[CBZ-L-Leu-L-Phe-L-(TR-Gln)]-E-Propenone

Using the procedure described in Example 3 for the preparation ofcompound 2, ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,1-pyrrolidin-1-yl-3-[BOC-L-(Tr-Gln)]-E-propenone (0.613 g, 1.166 mmol)was deprotected and coupled with CBZ-L-Leu-L-Phe (0.481 g, 1.166 mmol),yielding 1-pyrrolidin-1-yl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenone(0.668 g, 67%) as a white foam: R_(f)=0.5 (10% MeOH/CHCl₃); IR (thinfilm) 3294, 1702 cm⁻¹; ¹H NMR (CDCl₃) δ 0.84 (m, 6H), 1.31 (m, 1H), 1.46(m, 1H), 1.81-1.94 (m, 6H), 2.28 (m, 2H), 2.96 (m, 1H), 3.15 (m, 1H),3.39-3.50 (m, 4H), 3.95 (m, 2H), 4.87-5.11 (m, 4H), 6.14 (d, 1H, J=15.3Hz), 6.45 (d, 1H, J=7.8 Hz), 6.67 (dd, 1H, J=14.8, 4.8 Hz), 6.82 (d, 1H,J=8.1 Hz), 7.08-7.33 (m, 25H), 7.44 (d, 1H, J=8.1 Hz); MS (M+H) 862.

Preparation ofProduct-1-Pyrrolidin-1-yl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenone

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate,1-pyrrolidin-1-yl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenone (0.668 g,0.776 mmol) was deprotected to provide this final product (0.320 g, 67%)as a white solid: mp=195-196° C. (dec); R_(f)=0.4 (10% MeOH/CHCl₃); IR(thin film) 3289, 1684 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.79 (dd, 6H, J=12.1,6.5 Hz), 1.29 (m, 1H), 1.47 (m, 1H), 1.68-1.87 (m, 6H), 2.05 (m, 2H),2.84 (m, 1H), 3.01 (m, 1H), 3.29-3.40 (m, 4H), 3.94 (m, 1H), 4.44 (m,2H), 5.01 (m, 2H), 6.14 (d, 1H, J=14.9 Hz), 6.507 (dd, 1H, J=15.4, 5.8Hz), 6.76 (bs, 1H), 7.14-7.35 (m, 10H), 7.46 (d, 1H, J=7.8 Hz).7.95-8.02 (m, 2H); HRMS calcd for C₃₄H₄₅N₅O₆620.3448 (M+H), found620.3437; Anal. (C₃₄H₄₅N₅O₆0.2 Ch₂Cl₂) C, H, N.

Example 14 Preparation of Compound 22:N,N-Dimethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenamide Preparation ofIntermediate N,N-Dimethyl-3-[BOC-L-(Tr-Gln)]-E-Propenamide

Using the procedure described in Example 13 for the preparation of1-pyrrolidin-1-yl-3-[BOC-L-(Tr-Gln)]-E-propenone,3-[BOC-L-(Tr-Gln)]-E-propenone acid (1.05 g, 2.04 mmol) was coupled withN,N-dimethylamine (0.167 g, 2.04 mmol) to provide the amide (0.848 g,77%) as a white foam: R_(f)=0.6 (10% MeOH/CHCl₃); IR (thin film) 3297,1690 cm⁻¹; ¹H NMR (CDCl₃) δ 1.42 (s, 9H), 1.81 (m, 1H), 1.93 (m, 1H),2.38 (m, 2H), 2.98 (s, 3H), 3.03 (s, 3H), 4.27 (bs, 1H), 4.84 (d, 1H,J=7.2 Hz), 6.31 (dd, 1H, J=15.1, 1.4 Hz), 6.65 (dd, 1H, J=15.3, 5.9 Hz),6.94 (bs, 1H), 7.19-7.33 (m, 15H); Anal (C₃₂H₂₉N₃O₄0.9 CH₂CL₂), C, H, N.

Preparation of IntermediateN,N-Dimethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenamide

Using the procedure described in Example 3 for the preparation ofcompound 2, ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,N,N-dimethyl-3-[BOC-L-(Tr-Gln)]-E-propenamide (0.726 g, 1.567 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.646 g, 1.567 mmol) toprovide the product (0.417 g, 32%) as a white foam: R_(f)=0.5 (10%MeOH/CIICl₃); IR (thin film) 3291, 1702 cm⁻¹; ¹H NMR (CDCl₃) δ 0.84 (m,6H), 1.30 (m,1H), 1.47 (m, 1H), 1.74 (m, 1H), 1.94 (m, 3H), 2.56 (s,3H), 2.96 (m, 1H), 3.15 (m, 1H), 2.99 (d, 6H, J=13.4 Hz), 3.94 (m, 1H),4.54 (m, 2H), 4.87 (s, 2H), 5.00 (d, 2H, J=5.3 Hz), 6.28 (d, 1H, J=14.9Hz), 6.42 (d, 1H, J=7.8 Hz), 6.63 (dd, 1H, J=15.3, 5.0 Hz), 6.81 (d, 1H,J=8.4 Hz), 7.06 (bs 1H), 7.10-7.36 (m, 25H); Anal (C₅₁H₅₇N₅O₆3.0 H₂O) C,H, N.

Preparation of Product-N,N-Dimethyl-3-(CBZ-L-Phe-L-Gln)-E-Propenamide

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenoate,N,N-dimethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenamide (0.417 g, 0.5mmol) was deprotected to provideN,N-dimethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-propenamide (0.214 g, 72%) as awhite solid: mp=174-175° C. (dec); R_(f)=0.34 (MeOH/CHCl₃); IR (thinfilm) 3284, 1684 cm⁻¹; ¹ H NMR (DMSO-d₆) δ 0.79 (dd, 6H, J=12.1, 6.5Hz), 1.30 (m, 1H), 1.47 (m, 1H), 1.70 (m, 2H), 2.06 (m, 2H), 2.84 (m,1H), 2.98 (s, 3H), 3.03 (s, 3H), 3.94 (m, 1H), 4.44 (m, 2H), 4.95-5.07(m, 4H), 6.27 (d, 1H, J=15.3 Hz), 6.47 (dd, 1H, J=15.3, 5.6 Hz), 6.75(bs, 1H), 7.14-7.35 (m, 10H), 7.46 (d, 1H, J=7.5 Hz), 7.96-8.01 (m, 2H);HRMS calcd for C₃₂H₄₃N₅O₆ 594.3291 (M+H), found 594.3281. Anal.(C₃₂H₄₃N₅O₆·1.0 CH₂Cl₂) C, H, N.

Example 15 Preparation of Compound 24:1-Phenyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenone Preparation ofIntermediate 2-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Vinyl) Pyridine

2-Picolytriphenylphosphonium chloride/NaNH₂ (0.345 g, 0.76 mmol) wasdissolved in 10 mL of THF. CBZ-L-Leu-L-Phe-L-(Tr-Glutaminal) (0.53 g,0.69 mmol) was dissolved in 5 mL of THF and added dropwise to the yieldsolution at room temperature, which was allowed to stir overnight. Thesolvent was removed in vacuo, and the crude product purified by columnchromatography eluting with a gradient of 1-5% MeOH in CHCl₃ to give0.353 g (61%) of a white glassy solid: IR (KBr) 3295, 3061, 2953, 1952,1881, 1649, 1539, 1234, 1045, 972, 750, 696 cm⁻¹; ¹H NMR (DMSO-d₆) δ0.78 (t, 6H, J=7.0 Hz), 1.30 (m, 2H), 1.46 (m, 1H), 1.70 (m, 2H), 2.27(m, 2H), 2.78 (m, 1H), 3.03 (m, 1H), 3.97 (m, 1H), 4.42 (m, 1H), 4.52(m, 1H), 4.96 (d, 1H, J=12.0 Hz), 5.03 (d, 1H, J=12.0 Hz), 6.38 (d, 1H,J=16.0 Hz), 6.60 (dd, 1H, J=16.0, 6.0 Hz), 7.10-7.34 (m, 27H), 7.42 (d,1H, J=8.0 Hz), 7.73 (t, 1H, J=7.5 Hz), 7.92 (d, 1H, J=8.5 Hz), 8.07 (d,1H, J=8.5 Hz), 8.49 (d, 1H, J=5.0 Hz), 8.59 (s, 1H); MS (M+H) 842. Anal.(C₅₃H₅₅N₅O₅0.75 H₂O) C, H, N.

Preparation of Intermediate2-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Pyridine

Using the procedure described in Example 32 for the preparation ofcompound 20, diethyl-[2-(CBZ-L-Leu-L-Gln)-E-vinyl] phosphonate,2-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-vinyl] pyridine was synthesized from2-(CBZ-L-Leu-L-Phe-L-(Tr-Gln)-E-vinyl pyridine in 69% yield as a whitesolid: IR (KBr) 3291, 3059, 2955, 2359, 1694, 1641, 1539, 1234, 1119,1047, 970, 743, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.78 (m, 6H), 1.32 (m, 2H),1.49 (m, 1H), 1.77 (m, 2H), 2.11 (t, 2H, J=7.0 Hz), 2.86 (m, 1H), 3.01(m, 1H), 3.96 (m, 1H), 4.41 (m, 1H), 4.51 (m, 1H), 4.98 (d, 1H, J=13.0Hz), 5.04 (d, 1H, J=13.0 Hz), 6.39 (d, 1H, J=16.0 Hz), 6.60 (dd, 1H,J=16.0, 6.0 Hz), 6.75 (bs, 1H), 7.08-7.34 (m, 13H), 7.45 (d, 1H, J=8.0Hz), 7.73 (dt, 1H, J=7.5, 1.5 Hz), 7.97 (d, 1H, J=8.0 Hz), 8.07 (d, 1H,J=8.0 Hz), 8.50 (d, 1H, J=4.0 Hz); HRMS calcd for C₃₄H₄₁N₅O₅600.3186(M+H), found 600.3198. Anal. (C₃₄H₄₁N₅O₅1.0 H₂O) C, H, N.

Preparation of Intermediate1-Phenyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenone

Using the procedure described in Example 1 for the preparation ofcompound 12, ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate,this compound was synthesized from CBZ-L-Leu-L-Phe-L-Tr-glutaminal and(benzoylmethylene)triphenylphosphorane to give 0.38 g of crude material(impure with triphenylphosphine oxide), which was used without furtherpurification.

Preparation of Product-1-Phenyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenone

To 0.38 g of 1-phenyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenone, impurewith triphenylphosphine oxide, was added 10 mL of CH₂Cl₂. TFA (1mL) wasadded to this solution, and the reaction was stirred at room temperaturefor four hours. The reaction was poured into an EtOAc/saturated NaHCO₃solution and agitated until white solids began to precipitate out of theorganic layer. The aqueous layer was separated, and the solids filteredand washed with EtOAc to give compound 14 (0.0795 g, 20% yield from thealdehyde; 2 steps) as a white solid: IR (KBr) 3408, 3293, 3063, 2955,1653, 1539, 1449, 1283, 1234, 1121, 1047, 970, 698 cm⁻¹; ¹H NMR(DMSO-d₆) δ 0.78 (m, 6H), 1.31 (m, 2H), 1.45 (m, 1H), 1.76 (m, 2H), 2.11(t, 2H, J=8.0 Hz), 2.89 (m, 1H), 3.01 (m, 1H), 3.97 (m, 1H), 4.51 (m,2H), 4.97 (d, 1H, J=13.0 Hz), 5.05 (d, 1H, J=13.0 Hz), 6.76 (dd, 1H,J=15.0, 5.0 Hz), 6.77 (bs, 1H), 6.91 (d, 1H, J=15.0 Hz), 7.02-7.34 (m,11H), 7.47 (d, 1H, J=7.0 Hz), 7.54 (m, 2H), 7.66 (t, 1H, J=7.0 Hz), 7.93(d, 2H, J=7.0 Hz), 8.04 (d, 1H, J=8.0 Hz), 8.10 (d, 1H, J=8.5 Hz); HRMScalcd for C₃₆H₄₃N₄O₆627.3182 (M+H), found 627.3199. Anal. (C₃₆H₄₃N₄O₆)C, H, N.

Example 16 Preparation of Compound 26:Ethyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-Gln]-E-PropenoatePreparation of Intermediate BOC-L-(4-Cl-Phe)-L-(Tr-Glutaminol)

BOC-L-4-Cl-Phe (0.90 g, 3.0 mmol) was dissolved in 30 mL of THF.Carbonyldiimidazole (0.49 g, 3.0 mmol) was added, and the reaction wasallowed to stir at room temperature for one hour. L-(Tr-Glutaminol) (1.2g, 3 mmol) was added, and the reaction was stirred overnight at roomtemperature. The solvent was removed in vacuo, and the product waspurified by flash column chromatography eluting with 3% MeOH/CHCl₃ toyield 1.57 g (80%) of a white solid: IR (KBr) 3416, 3302, 3057, 3024,2978, 2934, 1663, 1491, 1447, 1366, 1250, 1165, 752, 700 cm⁻¹; ¹ H NMR(DMSO-d₆) δ 1.28 (s, 9H), 1.44 (m, 1H), 1.66 (m, 1H), 2.26 (m, 2H), 2.72(m, 1H), 2.91 (m, 1H), 3.18 (m, 2H), 3.64 (m, 1H), 4.07 (m, 1H), 4.67(t, 1H, J=5.0 Hz), 7.05-7.32 (m, 19H), 6.86 (d, 1H, J=8.5 Hz), 7.62 (d,1H, J=8.5 Hz), 8.48 (s, 1H). Anal. (C₃₈H₄₂N₃O₅Cl1.0 H₂O) C, H, N.

Preparation of Intermediate L-(4-Cl-Phe)-L-(Tr-Glutaminol)HydrochlorideSalt

BOC-L-(4-Cl-Phe)-L-(Tr-Glutaminol) (1.57 g, 2.4 mmol) was dissolved in aminimum amount of CH₂Cl₂ (˜5 mL) followed by 50 mL of Et₂O. AnhydrousHCl gas was bubbled into the solution until a white solid precipitatedfrom solution. The reaction was allowed to stir at room temperatureovernight, and the resulting solid was filtered and washed with Et₂O,giving 1.19 g (84%) of a white crystalline material: IR (KBr) 3246,3057, 3028, 2934, 1668, 1494, 1447, 1089, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ1.48 (m, 1H). 1.71 (m, 1H), 2.30 (m, 2H), 2.94-3.17 (m, 3H), 3.27 (m,1H), 3.67 (br, 2H), 3.98 (m, 1H), 7.07-7.40 (m, 19H), 8.28 (bs, 3H),8.34 (d, 1H, J=8.8 Hz), 8.54 (s, 1H). Anal. (C₃₃H₃₄N₃O₃Cl1.0 HC10.75H₂O) C, H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-(Tr-Glutaminol)

4-Methoxyindole-2-carboxylic acid (0.36 g, 1.87 mmol) was suspended in10 mL of CH₂Cl₂. To this suspension was added N-hydroxysuccinimide (0.23g, 1.97 mmol) and 2 mL of DMF to dissolve all solids.Dicyclohexylcarbodiimide (0.41 g, 1.97 mmol) was added and the reactionmixture was stirred at room temperature for 4 hours. At this time themixture was then filtered into a separate flask containing (1.17 g, 1.97mmol) of L-(4-Cl-Phe)-L-(Tr-glutaminol) HCl salt, 0.41 mL (2.95 mmol) ofEt₃N, 10 mL of CH₂Cl₂, and 2 mL of DMF, removing theN,N′-dicyclohexylurea precipitate. The reaction was allowed to stirovernight at room temperature. The solvents were removed in vacuo, andthe resulting crude product was purified by flash column chromatographyeluting with 3% (anhydrous NH₃/MeOH)/CHCl₃ to afford 0.53 g (39%) of awhite solid: IR (KBr) 3290, 3057, 2933, 1653, 1491, 1360, 1257, 1098,754, 698 cm¹; ¹H NMR (DMSO-d₆) δ 1.50 (m, 1H), 1.74 (m, 1H), 2.28 (m2H), 3.02 (m, 2H), 3.24 (m, 2H), 3.66 (m, 1H), 3.87 (s, 3H), 4.65 (m,1H), 4.70 (m, 1H), 6.49 (m, 1H, J=7.3 Hz), 6.94-7.38 (m, 22H), 7.86 (d,1H, J=8.8 Hz), 8.49 (d, 1H, J=8.8 Hz), 8.53 (s, 1H), 11.50 (s, 1H).Anal. (C₄₃H₄₁N₄O₅C10.75 H₂ O) C, H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-(Tr-Glutaminal)

N-(4-Methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminol) (1.13 g,1.55 mmol) was dissolved in 15 mL of DMSO. o-Iodoxybenzoic acid (1.30 g,4.66 mmol) was added to this solution, and dissolved after a few minutesof stirring at room temperature. After two hours the DMSO was removedunder reduced pressure. The residue was twice diluted with CH₂Cl₂, andthe solvent was evaporated to remove any residual DMSO. The residue wasdiluted with EtOAc, and the white precipitate was triturated andfiltered off.

The organic solvent was washed with 10% Na₂S₂O₃/10% NaHCO₃ solution,water, and brine before drying over Na₂SO₄. The solvent was removed togive 0.85 g (76%) of a white glassy solid which was used immediatelywithout further purification: ¹H NMR (DMSO-d₆) δ 1.72 (m, 2H), 2.32 (m,2H), 3.04 (m, 1H), 3.11 (m, 1H), 3.87 (m, 3H), 4.05 (m, 1H), 4.81 (m,1H), 6.49 (d, 1H, J=7.3 Hz), 6.94-7.39 (m, 22H), 8.60 (m, 2H), 8.63 (s,1H), 9.34 (s, 1H), 11.48 (s, 1H).

Preparation of IntermediateEthyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-(tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofcompound 12, ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate,this compound was synthesized fromN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminal) in 59%yield as a white solid: IR (KBr) 3302, 3057, 2934, 1958, 1896, 1659,1491, 1260, 1096, 1036, 833, 756, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.22 (t,3H, J=6.0 Hz), 1.72 (m, 2H), 2.24 (m, 2H), 3.05 (m, 2H), 3.88 (s, 3H),4.12 (q, 2H, J=6.0 Hz), 4.43 (m, 1H), 4.78 (m, 1H), 5.74 (d, 1H, J=14.0Hz), 6.50 (d, 1H, J=7.7 Hz), 6.77 (dd, 1H, J=16.0, 5.0 Hz), 6.93-7.57(m, 22H), 8.33 (d, 1H, J=7.7 Hz), 8.56 (d, 1H, J=7.7 Hz), 8.60 (s, 1H),11.51 (s, 1H). Anal. (C₄₇H₄₅N₄O₆Cl0.5 H₂O) C, H, N.

Preparation ofProduct-Ethyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 32 for the preparation ofcompound 20, diethyl-[2-(CBZ-L-Leu-L-PheL-Gln)-E-vinyl] phosphonate,this compound was synthesized by deprotection ofethyl-3-[N-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-Gln)]-E-propenoate.The product was purified by flash silica gel chromatography eluting with2-3% MeOH/CHCl₃ to give 0.16 g (73%) of an off-yellow solid: IR (KBr)3420, 3289, 2930, 2838, 1722, 1663, 1622, 1541, 1261, 1184, 1101, 976,754 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.21 (t, 3H), J=7.0 Hz), 1.74 (m, 2H), 2.11(t, 2H, J=8.0 Hz), 3.02 (m, 2H), 3.88 (s, 3H), 4.12 (q, 2H, J=7.0 Hz),4.42 (m, 1H), 4.68 (m, 1H), 5.74 (dd, 1H, J=16.0, 1.5 Hz), 6.47 (d, 1H,J=5.0 Hz), 6.75 (bs, 1H), 6.76-6.81 (m, 2H), 6.96 (d, 1H, J=8.5 Hz),7.07 (t, 1H, J=8.0 Hz), 7.24-7.38 (m, 5H, 8.33 (d, 1H, J=8.0 Hz), 8.58(d, 1H, J =8.5 Hz), 11.52 (s, 1H); HRMS calcd for C₂₈H₃₁N₄O₆Cl+Cs687.0986 (M+Cs), found 687.0976. Anal. (C₂₈H₃₁N₄O₆Cl) C, H, N.

Example 17 Preparation of Compound 27:Ethyl-3-]N-(4-Methoxyindole-2-Carbonyl)-L-(4-F-Phe)-L-Gln]-E-PropenoatePreparation of Intermediate BOC-L-(4-F-Phe)-L-(Tr-Glutaminol)

Using the procedure described in Example 16 for the preparation ofBOC-L-(4-Cl-Phe)-L-(Tr-glutaminol), this compound was synthesized fromBOC-L-4-F-Phe and L-(Tr-glutaminol) in 80% yield. White solid: IR (KBr)3416, 3308, 3057, 2978, 2932, 1663, 1510, 1368, 1223, 1167, 1051, 752,700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.28 (s, 9H), 1.44 (m, 1H), 1.68 (m, 1H),2.25 (m, 2H), 2.70 (m, 1H), 2.90 (m, 1H), 3.25 (m, 2H), 3.36 (m, 1H),4.10 (m, 1H), 4.67 (t, 1H, J=5.0 Hz), 7.04-7.28 (m, 19H), 6.85 (d, 1H,J=8.5 Hz), 7.61 (d, 1H, J=8.0 Hz), 8.48 (s, 1H). Anal. (C₃₈H₄₂N₃O₅F0.75H₂O) C, H, N.

Preparation of Intermediate L-(4-F-Phe)-L-(Tr-Glutaminol)HydrochlorideSalt

Using the procedure described in Example 16 for the preparation ofL-(4-Cl-Phe)-L-(Tr-glutaminol) hydrochloride salt, this salt wassynthesized from BOC-L-(4-F-Phe)-L-(Tr-glutaminol) in 79% yield. Whitecrystalline solid: IR (KBr) 3245, 3057, 2361, 1668, 1510, 1447, 1223,766, 700 cm ⁻¹; ¹H NMR (DMSO-d₆) δ 1.47 (m, 1H), 1.72 (m, 1H), 2.30 (m,2H), 2.94-3.16 (m, 3H), 3.23 (m, 1H), 3.65 (bs, 2H), 3.95 (m, 1H),7.09-7.32 (m, 19H), 8.28 (m, 4H), 8.54 (s, 1H), Anal. (C₃₃H₃₄N₃O₃F1.0HCl1.0 H₂O) C, H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(4-F-Phe)-L-(Tr-Glutaminol)

Using the procedure described in Example 16 for the preparation ofN-(4-methoxyindole-2-carbonyl-L-(4-Cl-Phe)-L-(Tr-glutaminol), thisintermediate was synthesized from 4-methoxyindole-2-carboxylic acid andL-(4-F-Phe)-L-(Tr-glutaminol) HCl salt, in 40% yield. White solid: IR(KBr) 3314, 3059, 2938, 1956, 1888, 1653, 1510, 1361, 1255, 1097, 835,756, 700 cm⁻¹; ¹H NMR (DMSO-d₆)δ 1.58 (m, 1H), 1.81 (m, 1H), 2.28 (m,2H), 3.02 (m, 2H), 3.23 (m, 2H), 3.67 (m, 1H), 3.87 (s, 3H), 4.69 (m,2H), 6.49 (m, 1H, J=7.3 Hz), 6.94-7.39 (m, 22H), 7.84(d, 1H, J=8.5 Hz),8.48 (d, 1H, J=8.5 Hz), 8.53 (s, 1H), 11.49 (s, 1H). Anal.(C₄₃H₄₁N₄O₅F1.0H₂O) C, H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(4-F-Phe)-L-(Tr-Glutaminal)

Using the oxidation procedure described in Example 16 for thepreparation ofN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminal), thisaldehyde was prepared in 80% yield fromN-(4-methoxyindole-2-carbonyl)-L-(4-F-Phe)-L-(Tr-glutaminol). Glassywhite solid: ¹H NMR (DMSO-d₆) δ 1.72 (m, 2H), 2.37 (m, 2H), 3.03 (m,1H), 3.17 (m, 1H), 3.87 (s, 3H), 4.09 (m, 1H), 4.47 (m, 1H), 6.49 (d,1H, J=7.7 Hz), 6.94-7.41 (m, 22H), 8.58 (m, 2H), 8.63 (s, 1H), 9.32 (s,1H), 11.49 (s, 1H).

Preparation of IntermediateEthyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(4-F-Phe)-L-(Tr-Gln)-]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofcompound 12, ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate,this vinyl ester was synthesized fromN-(4-methoxyindole-2-carbonyl)-L-(4-F-Phe)-L-(Tr-glutaminal) and(carbethoxymethylene)triphenyl-phosphorane in 60% yield. White solid: IR(KBr) 3300, 3061, 2938, 1958, 1890, 1653, 1510, 1368, 1260, 1100, 1036,835, 756, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.20 (t, 3H, J=7.0 Hz), 1.70 (m,2H), 2.35 (m, 2H), 3.01 (m, 2H), 3.87 (s, 3H), 4.11 (q, 2H, J=7.0 Hz),4.41 (m, 1H), 4.67 (m, 1H), 5.68 (d, 1H, J=16.0 Hz), 6.49 (d, 1H, J=7.7Hz), 6.74 (dd, 1H, J=16.0, 5.0 Hz), 6.97-7.38 (m, 22H), 8.31 (d, 1H,J=8.5 Hz), 8.55 (d, 1H, J=8.5 Hz), 8.58 (s, 1H), 11.51 (s, 1H). Anal.(C₄₇H₄₅N₄O₆F·1.0H₂O) C, H, N.

Preparation ofProduct-Ethyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(4-F-Phe)-L-Gln)]-E-Propenoate

Using the procedure described in Example 32 for the preparation ofcompound 20, diethyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-vinyl] phosphonate,this compound was synthesized by deprotection ofethyl-3-[N-(4-methoxyindole-2-carbonyl)-L-(4-F-Phe)-L-(Tr-Gln]-E-propenoatein 50% yield: White crystalline solid: IR (KBr) 3422, 3293, 2932, 1719,1665, 1620, 1541, 1510, 1369, 1261, 1182, 1101, 752 cm⁻¹; ¹H NMR(DMSO-d₆)δ 1.21 (t, 3H, J=7.0 Hz), 1.73 (m, 2H), 2.10 (t, 2H, J=8.0 Hz),3.02 (m, 2H), 3.88 (s, 3H), 4.13 (q, 2H, J=7.0 Hz), 4.43 (m, 1H), 4.67(m, 1H), 5.67 (dd, 1H, J=16.0, 1.5 Hz), 6.49 (d, 1H, J=7.0 Hz), 6.75(bs, 1H), 6.76 (dd, 1H, J=16.0, 5.5 Hz), 6.96 (d, 1H, J=8.5 Hz),7.03-7.10 (m, 3H), 7.23 (bs, 1H), 7.31-7.39 (m, 3H), 8.31 (d, 1H, J=8.0Hz), 8.57 (d, 1H, J=8.0 Hz), 11.51 (s, 1H); HRMS calcd forC₂₈H₃₁N₄O₆F+Cs 671.1282 (M+Cs), found 671.1288. Anal. (C₂₈H₃₁N₄O₆F) C,H, N.

Example 18 Preparation of Compound 28:Ethyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(3-F-Phe)-L-Gln]-E-PropenoatePreparation of Intermediate BOC-L-(3-F-Phe)-L-(Tr-glutaminol)

Using the procedure described in Example 16 for the preparation ofBOC-L-(4-Cl-Phe)-L-(Tr-glutaminol), this compound was synthesized fromBOC-L-3-F-Phe and L-(Tr-glutaminol) in 74% yield. White solid: IR (KBr)3410, 3302, 3059, 3030, 2974, 2934, 1663, 1491, 1448, 1250, 1167, 1051,752, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.28 (s, 9H), 1.46 (m, 1H), 1.17 (m,1H), 2.26 (m, 2H), 2.74 (m, 1H), 2.95 (m, 1H), 3.19 (m, 2H), 3.65 (m,1H), 4.11 (m, 1H), 4.67 (t, 1H, J=5.0 Hz), 6.97-7.32 (m, 19H), 6.89 (d,1H, J=8.5 Hz), 7.58 (d, 1H, J=8.5 Hz), 8.48 (s, 1H). Anal.(C₃₈H₄₂N₃O₅F1.0 H₂O) C, H, N.

Preparation of Intermediate L-(3-F-Phe)-L-(Tr-Glutaminol) HydrochlorideSalt

Using the procedure described in Example 16 for the preparation ofL-(4-Cl-Phe)-L-(Tr-glutaminol) hydrochloride salt, this salt wassynthesized from BOC-L-(3-F-Phe)-L-(Tr-glutaminol) in 88% yield. Whitecrystalline solid: IR (KBr) 3231, 3047, 1668, 1491, 1447, 1254, 1145,1036, 752, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.45 (m, 1H), 1.72 (m, 1H), 2.30(m, 2H), 2.96-3.11 (m, 3H), 3.25 (m, 1H), 3.70 (m, 1H), 4.03 (m, 1H),7.06-7.38 (m, 19H), 8.30 (bs, 4H), 8.54 (s, 1H). Anal.(C₃₃H₃₄N₃O₃F1.0HCl0.5 H₂O) C, H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(3-F-Phe)-L-(Tr-Glutaminol)

Using the procedure described in Example 16 for the preparation ofN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminol), thisintermediate was synthesized from 4-methoxyindole-2-carboxylic acid andL-(3-F-Phe)-L-(Tr-glutaminol) HCl salt, in 60% yield. White solid: IR(KBr) 3291, 3057, 2936, 1956, 1890, 1653, 1361, 1256, 1100, 754, 698cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.58 (m, 1H), 1.81 (m, 1H), 2.28 (m, 2H), 3.02(m, 2H), 3.28 (m, 2H), 3.70 (m, 1H), 3.87 (s, 3H), 4.68 (m, 2H), 6.49(m, 1H, J=7.7 Hz), 6.94-7.28 (m, 22H), 7.85 (d, 1H, J=8.5 Hz), 8.50 (d,1H, J=8.5 Hz), 8.54 (s, 1H), 11.50 (s, 1H). Anal. (C₄₃H₄₁N₄O₅F1.0H₂O) C,H, N.

Preparation of IntermediateN-(4-Methoxyindole-2-Carbonyl)-L-(3-F-Phe)-L-(Tr-Glutaminal)

Using the oxidation procedure described in Example 16 for thepreparation ofN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminal), thisaldehyde was prepared in 77% yield fromN-(4-methoxyindole-2-carbonyl)-L-(3-F-Phe)-L-(Tr-glutaminol and was usedimmediately. Glassy white solid: ¹H NMR (DMSO-d₆) δ 1.68 (m, 2H), 2.37(m, 2H), 3.04 (m, 1H), 3.18 (m, 1H), 3.87 (m, 3H), 4.05 (m, 1H), 4.81(m, 1H), 6.49 (d, 1H, J=7.7 Hz), 6.94-7.30 (m, 22H), 8.60 (m, 2H), 8.62(s, 1H), 9.33 (s, 1H), 11.48 (s, 1H).

Preparation of IntermediateEthyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(3-F-Phe)-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1for the preparation ofcompound 12, ethyl-3-[CBZ-L-Leu-Phe-L-Met(sulfoxide)-E-propenoate, thisvinyl ester was synthesized fromN-(4-methoxyindole-2-carbonyl)-L-(3-F-Phe)-L-(Tr-glutaminal) and(carbethoxymethylene)triphenyl-phosphorane in 68% yield. White solid: IR(KBr) 3293, 3057, 2934, 1956, 1894, 1657, 1491, 1368, 1260, 1100, 1036,978, 756, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.20 (t, 3H, J=7.0 Hz), 1.69 (m,2H), 2.25 (m, 2H), 3.02 (m, 2H), 3.87 (s, 3H), 4.11 (q, 2H, J=7.0 Hz),4.42 (m, 1H), 4.69 (m, 1H), 5.71 (d, 1H, J=16.0 Hz), 6.49 (d, 1H, J=8.0Hz), 6.75 (dd, 1H, J=16.0, 5.0 Hz), 6.91-7.29 (m, 22H), 8.32 (d, 1H,J=8.0 Hz), 8.56 (d, 1H, J=8.0 Hz), 8.59 (s, 1H), 11.51 (s, 1H). Anal.(C₄₇H₄₅N₄O₆F0.5 H₂O) C, H, N.

Preparation ofProduct-Ethyl-3-[N-(4-Methoxyindole-2-Carbonyl)-L-(3-F-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 32 for the preparation ofcompound 20, diethyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-vinyl]phosphonate,this compound was synthesized by deprotection ofethyl-3-[N-(4-methoxyindole-2-carbonyl)-L-(3-F-Phe)-L-(Tr-Gln)]-E-propenoatein 52% yield. White solid: IR (KBr) 3283, 2932, 1663, 1539, 1370, 1256,1188, 1098, 1036, 978, 752 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.21 (t, 3H, J=7.0Hz), 1.73 (m, 2H), 2.11 (t, 2H, J=7.0 Hz), 3.07 (m, 2H), 3.88 (s, 3H),4.11 (q , 2H, J=7.0 Hz), 4.49 (m, 1H), 4.75 (m, 1H), 5.72 (dd, 1H,J=16.0, 1.5 Hz), 6.49 (d, 1H, J=7.7 Hz), 6.80 (m, 2H), 6.98-7.31 (m,8H), 8.32 (d, 1H, J=8.0 Hz), 8.58 (d, 1H, J=8.0 Hz), 11.52 (s, 1H); HRMScalcd for C₂₈H₃₁N₄O₆F 539.2306 (M+H), found 539.2317. Anal.(C₂₈H₃₁N₄O₆F) C, H, N.

Example 19 Preparation of Compound 30:Ethyl-3-(CBZ-L-Phe-L-Gln)-E-Propenoate Preparation of IntermediateEthyl-3-[CBZ-L-Phe-L-(Tr-Gln)]-E-Propenoate

Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.60 g, 1.1 mmol), prepared as inExample 3, was deprotected and coupled with CBZ-L-Phe (0.31 g, 1.04mmol) using the procedure described in Example 28 for the preparation ofethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate to provideethyl-3-[CBZ-L-Phe-L-(Tr-Gln)]-E-propenoate (0.400 g, 53%) as a whitefoam: IR (thin film) 3298, 1651 cm⁻¹; ¹H NMR (CDCl₃) δ 1.21 (t, 3H,J=7.2 Hz), 1.65-1.75 (m, 1H), 1.90-1.93 (m, 1H), 2.29 (s, br, 1H),2.98-3.00 (m, 2H), 4.12 (q, 2H, J=7.2 Hz), 4.25-4.30 (m, 1H), 4.93 (d,1H, J=12.3 Hz), 4.50 (s, br, 1H), 5.01 (d, 1H, J=12.3 Hz), 5.23 (d, 1H,J=6.2 Hz), 5.63 (d, 1H, J=15.6 Hz), 6.39 (d, 1H, J=7.2 Hz), 6.61 (dd,1H, J=15.6, 5.6 Hz), 6.79 (s, 1H), 7.11-7.34 (m, 25H); Anal.(C₄₅H₄₅N₃O₆) C, H, N.

Preparation of Product-Ethyl-3-(CBZ-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, Ethyl-3-[CBZ-L-Phe-L-(Tr-Gln)]-E-propenoate (0.40 g, 0.58mmol) was deprotected to provide ethyl-3-(CBZ-L-Phe-L-Gln)-E-propenoate(0.15 g, 78%) as a white solid: mp=184-186° C.; IR (thin film) 3287,1637, 1533 cm⁻¹; ¹H NMR (DMSO-d₆)δ 1.21 (t, 3H, J=7.2 Hz), 1.64-1.80 (m,2H), 2.08 (t, 2H, J=7.6 Hz), 2.73-2.80 (m, 1H), 2.94 (dd, 1H, J=13.7,5.3 Hz), 4.11 (q, 2H, J=7.2 Hz), 4.20-4.26 (m, 1H), 4.28-4.39 (m, 1H),4.95 (s, 2H), 5.69 (d, 1H, J=15.9 Hz), 6.70 (d, 1H, J=5.3 Hz), 6.75-6.77(m, 2H), 7.17-7.35 (m, 11H), 7.53 (d, 1H, J=8.4 Hz), 8.20 (d, 1H, J=8.1Hz); Anal. (C₂₆H₃₁N₃O₅) C, H, N.

Example 20 Preparation of Compound 31:Ethyl-3-[N-(Propylsulfonyl)-L-Phe-L-Gln]-E-Propenoate Preparation ofIntermediate Ethyl-3-[BOC-L-Phe-L-(Tr-Gln)]-E-Propenoate

Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenoate (2.26 g, 4.16 mmol), prepared asin Example 3, was dissolved in 1,4-dioxane (15 mL). A solution of HCl in1,4-dioxane (4.0 M, 15 mL) was added dropwise. The reaction solution wasstirred at room temperature for 2 hours, then poured into a solution ofaqueous NaOH (1 M, 80 mL) in saturated aqueous NaHCO₃ (120 mL). Theresulting mixture was extracted with CH₂Cl₂ (2×200 mL). The combinedorganic phases were dried over Na₂SO₄ and concentrated to give the freeamine intermediate as a slightly yellow solid, which was used withoutfurther purification. This crude amine, BOC-L-Phe (1.10 g, 4.15 mmol)and 1-hydroxybenzotriazole hydrate (0.843 g, 6.24 mmol) were stirred indry CH₂Cl₂ (35 mL) under argon at room temperature. 4-Methylmorpholine(1.83 mL, 16.6 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (1.20 g, 6.26 mmol) were added sequentially. Afterstirring for 3.5 hours, the reaction mixture was poured into water (100mL), and the mixture was extracted with CH₂Cl₂ (2×100 mL). The combinedorganic phases were dried over Na₂SO₄ and concentrated. The residue waspurified by column chromatography (33% acetone in hexanes) to give theproduct (1.94 g, 68%) as a white foam: IR (thin film) 3413, 3310, 1708,1660 cm⁻¹; ¹H NMR (CDCl₃) δ 1.30 (t, 3H, J=7.2 Hz), 1.39 (s, 9H),1.64-1.77 (m, 1H), 1.88-2.00 (m, 1H), 225-2.31 (m, 2H), 2.94-3.07 (m,2H), 4.18 (q, 2H, J=7.2 Hz). 4.49-4.59 (m, 1H), 4.95 (bs, 1H), 5.66 (d,1H, J=15.9 Hz), 6.29 (m, 1H), 6.64 (dd, 1H, J=15.9, 5.3 Hz), 6.81 (bs,1H), 7.14-7.34 (m, 21H); Anal. (C₄₂H₄₇N₃O₆) C, H, N.

Preparation of Intermediate Ethyl-3-[L-Phe-L-(Tr-Gln)]-E-Propenoate

Ethyl-3-[BOC-L-Phe-L-(Tr-Gln)]-E-propenoate (0.300 g, 0.435 mmol) wasdissolved in 1,4-dioxane (2mL). A solution of HCl in 1,4-dioxane (4.0 M,2 mL) was added dropwise. The reaction solution was stirred at roomtemperature for 2.5 hours, then poured into a solution of aqueous NaOH(1 M, 10 mL) in saturated aqueous NaHCO₃ (20 mL). The resulting mixturewas extracted with Ch₂Cl₂ (3×40 mL). The combined organic phases weredried over Na₂SO₄ and concentrated to give the product as a foam (0.257g, quantitative) which was used without further purification.

Preparation of IntermediateEthyl-3-[N-(Propylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-Propenoate

Ethyl-3-[L-Phe-L-(Tr-Gln)-]E-propenoate was dissolved in dry CH₂Cl₂ (7mL) under argon and cooled to 0° C. NEt₃ (0.067 mL, 0.48 mmol) and1-propanesulfonyl chloride (0.054 mL, 0.48 mmol) were addedsequentially. After stirring for 1 hour, the reaction mixture wasallowed to warm to room temperature. More NEt₃ (0.100 mL, 0.714 mmol)and 1-propanesulfonyl chloride (0.086 mL, 0.76 mmol) were added. After1.5 hours more, the solvent was evaporated and the residue was purifiedby column chromatography (50% EtOAc in hexanes) to give the product as afoam (0.121 g, 40%): IR (thin film) 3292, 1713, 1652, 1312, 1144 cm⁻¹;¹H NMR (CDCl₃) δ 0.80 (t, 3H, J=7.5 Hz), 1.28 (t, 3H, J=7.2 Hz),1.34-1.58 (m, 2H), 1.67-1.81 (m, 1H), 1.92-2.04 (m, 1H), 2.32-2.56 (m,4H), 2.79 (dd, 1H, J=13.9, 8.9 Hz), 3.05 (dd, 1H, J=13.9, 5.5 Hz),3.96-4.05 (m, 1H), 4.17 (q, 2H, J=7.2 Hz), 4.49-4.59 (m, 1H), 5.14 (d,1H, J=8.7 Hz), 5.75 (dd, 1H, J=15.9, 1.7 Hz), 6.72 (dd, 1H, J=15.9, 5.3Hz), 6.94 (s, 1H), 7.02 (d, 1H, J=8.1 Hz), 7.12-7.33 (m, 20H); HRMS(M+Cs) calcd for C₄₀H₄₅N₃O₆S 828.2083, found 828.2063.

Preparation ofProduct—Ethyl-3-[N-(Propylsulfonyl)-L-Phe-L-Gln]-E-Propenoate

Ethyl-3-[N-(propylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate (0.100 g,0.143 mmol) was dissolved in CH₂Cl₂/TFA 1:1 (4 mL) under argon. Thebright yellow solution was stirred at room temperature for 30 minutes.CCl₄ (4 mL) was added and the solution was concentrated to dryness. Theresidue was triturated with Et₂O (3 mL) to give a white precipitatewhich was collected by filtration and washed with Et₂O (2×2 mL) to givethe product (0.048 g, 74%): mp=161-162° C.; IR (KBr) 3284, 3213, 1708,1666, 1543, 1314, 1138 cm⁻¹; ¹H NMR (acetone-d₆) δ 0.83 (t, 3H, J=7.5Hz), 1.25 (t, 3H, J=7.2 Hz), 1.39-1.62 (m, 2H), 1.73-2.02 (m, 2H),2.23-2.30 (m, 2H), 2.54-2.72 (m, 2H), 2.92 (dd, 1H, J=13.5, 8.9 Hz),3.15 (dd, 1H, J=13.5, 6.1 Hz), 4.14 (q, 2H, J=7.2 Hz), 4.12-4.21 (m,1H), 4.53-4.63 (m, 1H), 5.79 (dd, 1H, J=15.7, 1.7 Hz), 6.18 (bs, 1H),6.30 (d, 1H, J=8.7 Hz), 6.78 (dd, 1H, J=15.7, 5.4 Hz), 6.75 (bs, 1H),7.19-7.35 (m, 5H), 7.59 (d, 1H, J=8.1 Hz); Anal. (C₂₁H₃₁N₃O₆S) C, H, N.

Example 21 Preparation of Compound 32:Ethyl-3-[N-(Benzylsulfonyl)-L-Phe-L-Gln]-E-Propenoate Preparation ofIntermediate Ethyl-3-[N-(Benzylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-Propenoate

Ethyl-3-[L-Phe-L-(Tr-Gln)]-E-propenoate (0.250 g, 0.424 mmol) wasdissolved in dry CH₂Cl₂ (7 mL) under argon and cooled to 0° C.Triethylamine (0.118 mL, 0.847 mmol) and α-toluenesulfonyl chloride(0.162 g, 0.850 mmol) were added sequentially. After stirring for 45min, the solvent was evaporated and the residue was purified by columnchromatography (47% EtOAc in hexanes) to give the product as a whitefoam (0.154 g, 49%): IR (thin film) 3296, 1708, 1663, 1316, 1154 cm⁻¹;¹H NMR (CDCl₃) δ 1.29 (t, 3H, J=7.2 Hz), 1.59-1.72 (m, 1H), 1.91-2.03(m, 1H), 2.31-2.37 (m, 2H), 2.82 (dd, 1H, J=13.7, 7.2 Hz), 2.92 (dd, 1H,J=13.7, 7.2 Hz), 3.78-3.87 (m, 1H), 3.90 (d, 1H, J=13.9 Hz), 3.97 (d,1H, J=13.9 Hz), 4.17 (q, 2H, J=7.2 Hz), 4.44-4.54 (m, 1H), 4.96 (d, 1H,J=7.8 Hz), 5.59 (dd, 1H, J=15.7, 1.7 Hz), 6.51 (d, 1H, J=7.5 Hz), 6.63(dd, 1H, J=15.7, 5.1 Hz), 6.91 (s, 1H), 7.03-7.07 (m, 2H), 7.17-7.40 (m,23H); Anal. (C₄₄H₄₅N₃O₆S) C, H, N.

Preparation ofProduct—Ethyl-3-[N-(Benzylsulfonyl)-L-Phe-L-Gln]-E-Propenoate

This compound was prepared in 72% yield fromethyl-3-[N-(benzylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate using theprocedure described in Example 20 for the preparation ofethyl-3-[N-(propylsulfonyl)-L-Phe-L-Gln]-E-propenoate: mp=165-167° C.;IR (KBr) 3330, 3201, 1713, 1660, 1314 cm⁻¹; ¹H NMR (acetone-d₆) δ 1.25(t, 3H, J=7.2 Hz), 1.72-1.99 (m, 2H), 2.22-2.30 (m, 2H), 2.96 (dd, 1H,J=13.5, 7.3 Hz), 3.10 (dd, 1H, J=13.5, 7.0 Hz), 4.03-4.22 (m, 5H),4.51-4.62 (m, 1H), 5.72 (dd, 1H, J=15.6, 1.6 Hz), 6.18 (bs, 1H), 6.33(d, 1H, J=8.4 Hz), 6.72 (bs, 1H), 6.73 (dd, 1H, J=15.6, 5.4 Hz),7.19-7.35 (m, 10H), 7.55 (d, 1H, J=8.1 Hz); Anal. (C₂₅H₃₁N₃O₆S) C, H, N.

Example 22 Preparation of Compound 33:Ethyl-3-[N-(Ethylsulfonyl)-L-Phe-L-Gln]-E-Propenoate Preparation ofIntermediate Ethyl-3-[N-(Ethylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-Propenoate

This compound was prepared in 46% yield fromethyl-3-[L-Phe-L-(Tr-Gln)]-E-propenoate and ethanesulfonyl chlorideusing the procedure described in Example 21 for the preparation ofethyl-3-[N-(benzylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate. The materialwas purified by flash column chromatography (50% EtOAc in hexanes): IR(thin film) 3295, 1713, 1666, 1314, 1143 cm⁻¹; ¹H NMR (CDCl₃) δ 1.04 (t,3H, J=7.5 Hz), 1.29 (t, 3H, J=7.2 Hz), 1.68-1.81 (m, 1H), 1.95-2.06 (m,1H), 2.33-2.43 (m, 2H), 2.45-2.58 (m, 1H), 2.59-2.72 (m, 1H), 2.86 (dd,1H, J=13.7, 8.4 Hz), 3.09 (dd, 1H, J=13.7, 5.6 Hz), 3.96-4.04 (m, 1H),4.19 (q, 2H, J=7.2 Hz), 4.50-4.59 (m, 1H), 4.91 (bs, 1H), 5.72 (dd, 1H,J=15.9, 1.9 Hz), 6.71 (dd, 1H, J=15.9, 5.3 Hz), 6.87 (s, 1H), 6.96 (d,1H, J=7.8 Hz), 7.13-7.34 (m, 20H); Anal. (C₃₉H₄₃N₃O₆S) C, H, N.

Preparation of ProductEthyl-3-[N-(Ethylsulfonyl)-L-Phe-L-Gln]-E-Propenoate

This compound was prepared in 82% yield fromethyl-3-[N-(ethylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate using theprocedure described in Example 20 for the preparation of compound 31,ethyl-3-[N-(propylsulfonyl)-L-Phe-L-Gln]-E-propenoate: mp=150-151° C.;IR (KBr) 3284, 3225, 1713, 1655, 1314, 1138 cm⁻¹; ¹H NMR (acetone-d₆) δ1.05 (t, 3H, J=7.3 Hz), 1.26 (t, 3H, J=7.2 Hz), 1.74-1.87 (m, 1H),1.90-2.02 (m, 1H), 2.22-2.33 (m, 2H), 2.62-2.84 (m, 2H), 2.95 (dd, 1H,J=13.7, 8.7 Hz), 3.15 (dd, 1H, J=13.7, 6.2 Hz), 4.16 (q, 2H, J=7.2 Hz),4.13-4.23 (m, 1H), 4.54-4.64 (m, 1H), 5.78 (dd, 1H, J=15.9, 1.6 Hz),6.22 (bs, 1H), 6.34 (d, 1H, J=9.0 Hz), 6.78 (bs, 1H), 6.78 (dd, 1H,J=15.9, 5.6 Hz), 7.21-7.35 (m, 5H), 7.61 (d, 1H, J=8.1 Hz); Anal.(C₂₀H₂₉N₃O₆S) C, H, N.

Example 23 Preparation of Compound 34:Ethyl-3-[N-(Phenylsulfonyl)-L-Phe-L-Gln]-E-Propenoate Preparation ofIntermediate Ethyl-3-[N-(Phenylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-Propenoate

This compound was prepared in 55% yield fromethyl-3-[L-Phe-L-(Tr-Gln)]-E-propenoate and benzenesulfonyl chlorideusing the procedure described in Example 21 for the preparation ofethyl-3-[N-(benzylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate. The materialwas purified by flash column chromatography (47% EtOAc in hexanes): IR(thin film) 3295, 1713, 1660, 1308, 1161 cm⁻¹; ¹H NMR (CDCl₃) δ 1.29 (t,3H, J=7.2 Hz), 1.59-1.72 (m, 1H), 1.83-1.95 (m, 1H), 2.12-2.33 (m, 2H),2.82-2.94 (m, 2H), 3.82-3.91 (m, 1H), 4.18 (q, 2H, J=7.2 Hz), 4.31-4.41(m, 1H), 5.05 (d, 1H, J=7.8 Hz), 5.67 (dd, 1H, J=15.7, 1.7 Hz), 6.60(dd, 1H, J=15.7, 5.4 Hz), 6.72 (d, 1H, J=7.8 Hz), 6.79 (s, 1H),6.91-6.97 (m, 2H), 7.13-7.40 (m, 20H), 7.48-7.54 (m, 1H), 7.58-7.62 (m,2H); Anal. (C₄₃H₄₃N₃O₆S) C, H, N.

Preparation of ProdcutEthyl-3-[N-(Phenylsulfonyl)-L-Phe-L-Gln]-E-Propenoate

This compound was prepared in 83% yield fromethyl-3-[N-(phenylsulfonyl)-L-Phe-L-(Tr-Gln)]-E-propenoate using theprocedure described in Example 20 for the preparation ofethyl-3-[N-(propylsulfonyl)-L-Phe-L-Gln]-E-propenoate: mp=173-175° C.;IR (KBr) 3284, 3201, 1708, 1660, 1314, 1161 cm⁻¹; ¹H NMR (acetone-d₆) δ1.24 (t, 3H, J=7.2 Hz), 1.59-1.85 (m, 2H), 2.07-2.19 (m, 2H), 2.85 (dd,1H, J=13.5, 7.6 Hz), 2.99 (dd, 1H, J=13.5, 6.7 Hz), 4.03-4.16 (m, 1H),4.13 (q, 2H, J=7.2 Hz), 4.30-4.40 (m, 1H), 5.65 (dd, 1H, J=15.7, 1.6Hz), 6.21 (bs, 1H), 6.63 (dd, 1H, J=15.7, 5.6 Hz), 6.74 (bs, 1H), 6.75(d, 1H, J=8.7 Hz), 7.07-7.29 (m, 5H), 7.42-7.61 (m, 4H), 7.67-7.80 (m,2H); Anal. (C₂₄H₂₉N₃O₆S) C, H, N.

Example 24 Preparation of Compound 35:Ethyl-3-[CBZ-L-Leu-L-(4-F-Phe)-L-Gln]-E-Propenoate Preparation ofIntermediate CBZ-L-Leu-L-(4-F-Phe)-L-(Tr-Glutaminol)

Using the procedure described in Example 16 for the preparation ofN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminol), thisintermediate was synthesized from CBZ-L-Leu and the free base ofL-(4-F-Phe)-L-(Tr-glutaminol).HCl, in 68% yield as a white solid: IR(KBr) 3304, 3063, 2955, 1651, 1510, 1223, 1038, 752, 698 cm⁻¹; ¹H NMR(DMSO-d₆) δ 0.79 (m, 6H), 1.34 (m, 2H), 1.46 (m, 2H), 1.72 (m, 1H), 2.25(m, 2H), 2.80 (m, 1H), 2.99 (m, 1H), 3.16 (m, 1H), 3.26 (m, 1H), 3.64(m, 1H), 3.95 (m, 1H), 4.47 (m, 1H), 4.66 (t, 1H, J=5.5 Hz), 4.97 (d,1H, J=12.5 Hz), 5.02 (d, 1H, J=12.5 Hz), 7.01 (t, 2H, J=8.8 Hz),7.15-7.37 (m, 22H), 7.42 (d, 1H, J=7.7 Hz), 7.69 (d, 1H, J=8.5 Hz), 7.87(d, 1H, J=8 Hz), 8.54 (s, 1H).

Preparation of Intermediate CBZ-L-Leu-L-(4-F-Phe)-L-(Tr-Glutaminal)

Using the oxidation procedure described in Example 16 for thepreparation ofN-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-glutaminol), thisaldehyde was prepared from CBZ-L-Leu-L-(4-F-Phe)-L-(Tr-glutaminol) in92% yield as a white glassy solid, which was used immediately withoutfurther purification.

Preparation of IntermediateEthyl-3-[CBZ-L-Leu-L-(4-F-Phe)-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofcompound 12, ethyl-3-[CBZ-L-Leu-L-Phe-L-Met(sulfoxide)-E-propenoate,(carbethoxymethylene)triphenyl-phosphorane andCBZ-L-Leu-L-(4-F-Phe)-L-(Tr-glutaminal) were stirred together in THFgiving 0.37 g of the crude material contaminated with triphenylphosphineoxide which was subsequently used without further purification. A smallamount (27 mg) was purified by flash column chromatography (MeOH/CHCl₃)for spectral analysis: ¹H NMR (DMSO-d₆) δ 0.79 (t, 6H, J=7.0 Hz), 1.20(t, 3H, J=7.0 Hz), 1.23-1.82 (m, 5H), 2.25 (m, 2H), 2.85 (m, 1H), 2.95(m, 1H), 3.96 (m, 1H), 4.10 (q, 2H, J=7.0 Hz), 4.34 (m, 1H), 4.48 (m,1H), 4.96 (d, 1H, J=13.0 Hz), 5.02 (d, 1H, J=13.0 Hz), 5.57 (d, 1H,J=15.0 Hz), 6.67 (dd, 1H, J=15.0, 5.5 Hz), 7.01 (t, 2H, J=9.0 Hz),7.13-7.32 (m, 22H), 7.39 (d, 1H, J=8.0 Hz), 7.99 (d, 1H, J=8.0 Hz), 8.07(d, 1H, J=8.0 Hz), 8.58 (s, 1H).

Preparation of ProductEthyl-3-[CBZ-L-Leu-L-(4-F-Phe)-L-Gln)-E-Propenoate

This compound was prepared by the deprotection ofethyl-3-[CBZ-L-Leu-L-(4-F-Phe)-L-(Tr-Gln)]-E-propenoate using theprocedure describe in Example 32 for the preparation of compound 20, butin the absence of triisopropylsilane. The product was isolated as awhite solid in 58% yield (2 steps fromCBZ-L-Leu-L-(4-F-Phe)-L-(Tr-glutaminal). IR (KBr) 3439, 3293, 3067,2961, 1692, 1643, 1539, 1227, 1045, 984, 835, 698 cm⁻¹; ¹H NMR (DMSO-d₆)δ 0.80 (m, 6H), 1.21 (t, 3H, J=7.0 Hz), 1.26 (m, 2H), 1.45 (m, 1H), 1.71(m, 2H), 2.06 (t, 2H, J=7.5 Hz), 2.81 (m, 1H), 2.94 (m, 1H), 3.97 (m,1H), 4.10 (q, 2H, J=7.0 Hz), 4.37 (m, 1H), 4.47 (m, 1H), 4.98 (d, 1H,J=12.5 Hz), 5.04 (d, 1H, J=12.5 Hz), 5.59 (d, 1H, J=16.0 Hz), 6.68 (dd,1H, J=16.0, 5.5 Hz), 6.76 (bs, 1H), 7.01 (t, 2H, J=8.8 Hz), 7.19-7.34(m, 8H), 7.43 (d, 1H, J=8.0 Hz), 8.05 (m, 2H); HRMS calcd forC₃₂H₄₁N₄O₇F+Cs 745.2014 (M+Cs), found 745.2040 Anal. (C₃₂H₄₁N₄O₇F.1.25H₂O) C, H, N.

Example 25 Preparation of Compound 15:3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoic Acid Preparation of Intermediatetert-Butyl-3-[CBZ-L-Phe-L-(Tr-Gln)]-E-Propenoate

To 0.20 g (0.261 mmol) of CBZ-L-Leu-L-Phe-L-(Tr-glutaminal) was added 3mL of dry THF. To this stirred solution was added(tert-butoxycarbonylmethylene) triphenylphosphorane (0.098 g, 0.261mmol). The reaction mixture was stirred at room temperature overnight.The solvent was removed in vacuo, and the residue was subjected tocolumn chromatography with hexanes:EtOAc (6.5:3.5). The product wasobtained in 69% yield as a white foam.

Preparation of Product 3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoic Acid

tert-Butyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.157 g, 0.181mmol) was dissolved in an excess of TFA, and 10 drops of water wereadded. The mixture was stirred at room temperature for 1 hour andevaporated to dryness. CCl₄ was added and the mixture was concentratedin vacuo to azeotrope any remaining water. The residue was slurried inEt₂O and the resulting white solid was filtered and dried to give 0.053g (52%). mp=219-220° C. (dec); IR (thin film); 2949, 1690, 3269, 1639cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.80 (dd, 6H, J=9.0, 6.5 Hz), 1.23-1.38 (m,2H), 1.41-1.56 (m, 1H), 1.61-1.79 (m, 2H), 2.0-2.1 (m, 2H), 2.84 (dd,1H, J=13.6, 8.9 Hz), 2.99 (dd, 1H, J=13.5, 5.1 Hz), 3.91 (m, 1H),4.32-4.41 (m, 1H), 4.44-4.54 (m, 1H), 5.01 (dd, 1H, J=12.5, 12.1 Hz),5.64 (d, 1H, J=15.6 Hz), 6.64 (dd, 1H, J=15.6, 5.6 Hz), 6.76 (bs, 1H),7.14-7.38 (m, 11H), 7.43 (d, 1H, J=7.5 Hz), 7.97 (d, 1H, J=8.1 Hz), 8.04(d, 1H, J=8.1 Hz), 12.28 (bs, 1H).

Example 26 Preparation of Compound 14:3-(CBZ-L-Leu-L-Phe-DL-Gln)-E-Propenonitrile Preparation of Intermediate3-[BOC-DL-(Tr-Gln)]-E-Propenonitrile

A solution of diethyl cyanomethylphosphonate (0.202 mL, 1.25 mmol) indry THF (25 mL) was cooled to −78° C. After dropwise addition of asolution of sodium bis(trimethylsilyl)amide in THF (1.0 M, 1.25 mL), thereaction solution was stirred for 20 minutes. A solution ofBOC-L-(Tr-glutaminol) (0.590 g, 1.25 mmol) in dry THF (5 mL) was addeddropwise, and, after stirring 50 minutes more, saturated aqueous NH₄Cl(4 mL) was added. The reaction mixture was allowed to warm to roomtemperature, and the THF was evaporated. Water (10 mL) was added to theresidue, which was then extracted with CH₂Cl₂ (3×30 mL). The combinedorganic phases were dried over Na₂SO₄ and concentrated. The residue waspurified by flash column chromatography (38% EtOAc in hexanes) to givethe product (0.407 g, 66%) as a white foam: IR (thin film) 3321, 2225,1694, 1515 cm⁻¹; ¹H NMR (CDCl₃) δ 1.42 (s, 9H), 1.67-1.81 (m, 1H),1.82-1.97 (m, 1H), 2.34-2.42 (m, 2H), 4.23 (bs, 1H), 4.97-5.06 (m, 1H),5.39 (dd, 1H, J=16.3, 1.6 Hz), 6.56 (dd, 1H, J=16.3, 5.3 Hz), 6.77 (bs,1H), 7.15-7.33 (m, 15H).

Preparation of Intermediate (CBZ-L-Leu-L-Phe)₂O

CBZ-L-Leu-L-Phe (1.5 g, 3.6 mmol) was dissolved in dry CH₂Cl₂ (25 mL) atroom temperature under argon.1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.697 g,3.64 mmol) was added. The reaction solution was stirred for 20 hours,then diluted with CH₂Cl₂ (20 mL) and washed with water (2×20 mL). Thecombined organic phases were dried over Na₂SO₄ and concentrated to givethe anhydride product as a white semi-solid residue (1.18 g, 80%), whichwas used immediately in the next step of the reaction without furtherpurification or analysis.

Preparation of Intermediate3-[CBZ-L-Leu-Phe-DL-(Tr-Gln)]-E-Propenonitrile

3-[BOC-DL-(Tr-Gln)]-E-Propenonitrile (0.349 g, 0.704 mmol) was stirredin 2-propanol (9 mL) at room temperature. Perchloric acid (60%, 3.2 mL)was added dropwise. The resulting solution was stirred for 1 hour underan argon balloon, diluted with CH₂Cl₂ (100 mL), and poured into asolution of aqueous 1N NaOH/aqueous saturated NaHCO₃ (40 mL:70 mL). Thephases were mixed and separated. The aqueous phase was washed again withCH₂Cl₂ (2×100 mL). The combined organic phases were dried over Na₂SO₄and then concentrated to give the crude amine as a white solid (0.314g), which was used without further purification. This amine wasdissolved in acetone (15 mL) and added to the crude (CBZ-L-Leu-L-Phe)₂O(1.18 g, 1.46 mmol) in a round bottom flask. The reaction solution wasstirred at room temperature under an argon balloon. After stirring for4.5 hours, the solvent was evaporated, and the residue was purified byflash column chromatography (30% EtOAc in hexanes, then 30% acetone inhexanes) to give the product (0.448 g, 81%) as a white foam: IR (thinfilm) 3298, 2226, 1672, 1519 cm⁻¹; Anal. (C₄₉H₅₁N₅O₅) C, H, N.

Preparation of Product 3-(CBZ-L-Leu-L-Phe-DL-Gln)-E-Propenonitrile

3-[CBZ-L-Leu-L-Phe-DL-(Tr-Gln)]-E-Propenonitrile (0.381 g, 0.482 mmol)was dissolved in 1:1 CH₂Cl₂/TFA (14 mL) under argon, giving a brightyellow solution. After stirring for 30 minutes, the solvent wasevaporated. CCl₄ (15 mL) was added, and the resulting solution wasconcentrated (3 times). The residue was triturated with Et₂O (8 mL) togive a white solid, which was collected by filtration. This solid wasthen stirred in acetonitrile (4 mL), collected by filtration, washedwith acetonitrile (4 mL), washed with Et₂O (6 mL), and dried invacuo-(0.099 g, 38%): mp=178-184° C.; IR (KBr) 3401, 3284, 2225, 1689,1650, 1537 cm⁻¹; ¹H NMR (DMSO-d₆) (2 diastereomers) δ 0.69 (d, 3H, J=5.3Hz), 0.73 (d, 3H, J=5.1 Hz), 0.80 (d, 3H, J=6.6 Hz), 0.83 (d, 3H, J=6.6Hz), 1.10-1.20 (m, 3H, 1.26-1.40 (m, 2H), 1.46-1.85 (m, 5H), 1.99-2.09(m, 4H), 2.76 (dd, 1H, J=13.4, 10.9 Hz), 2.83-2.99 (m, 2H), 3.10 (dd,1H, J=13.6, 4.3 Hz), 3.85-3.93 (m, 1H), 3.96-4.05 (m, 1H), 4.28-4.52 (m,4H), 4.90-5.07 (m, 5H), 5.71 (d, 1H, J=16.4 Hz), 6.68 (dd, 1H, J=16.4,4.6 Hz), 6.78 (s, 2H), 6.88 (dd, 1H, J=16.3, 4.7 Hz), 7.16-7.37 (m,22H), 7.41-7.47 (m, 2H), 7.96 (d, 1H, J=8.2 Hz), 8.03-8.10 (m, 2H), 8.38(d, 1H, J=8.2 Hz); Anal. (C₃₀H₃₇N₅O₅) C, H, N.

Example 27 Preparation of Compound 6:N-Ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenamide Preparation ofIntermediate N-Ethyl-3-[BOC-L-(Tr-Gln)]-E-Propenamide

Isobutyl chloroformate (0.161 mL, 1.24 mmol) was added to a solution of3-[BOC-L-(Tr-Gln)]-E-propenoic acid (0.639 g, 1.24 mmol) and4-methylmorpholine (1.36 mL, 12.4 mmol) in CH₂Cl₂ at 0° C. The resultingsolution was stirred for 20 minutes at 0° C., then ethylaminehydrochloride (0.810 g, 9.93 mmol) was added. The reaction mixture waswarmed to 23° C. and was stirred for 24 hours, then was partitionedbetween water (100 mL) and a 9:1 mixture of CH₂Cl₂ and CH₃OH (2×100 mL).The organic layers were dried over Na₂SO₄ and were concentrated.Purification of the residue by flash column chromatography (5%CH₃OH/CH₂Cl₂) provided an oil, which was triturated with EtOAc to afforda white solid. The solid was filtered, washed with EtOAc (2×20 mL), andwas air-dried to give N-ethyl-3-[BOC-L-(Tr-Gln)]-E-propenamide (0.055 g,8%): mp=240° C. (dec); IR (thin film) 3255, 3085, 1715, 1665, 1612, 1529cm⁻¹; ¹H NMR (CDCl₃) δ 1.15 (t, 3H, J=7.2 Hz), 1.42 (s, 9H), 1.63-1.80(m, 1H), 1.83-2.05 (m, 1H), 2.34-2.39 (m, 2H), 3.29-3.38 (m, 2H), 4.26(s, br, 1H), 4.75 (s, br, 1H), 5.43 (s, br, 1H), 5.81 (d, 1H, J=15.4Hz), 6.65 (dd, 1H, J=15.4, 5.9 Hz), 6.85 (s, 1H), 7.19-7.33 (m, 15H);Anal. (C₃₃H₃₇N₃O₄) C, H, N.

Preparation of IntermediateN-Ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenamide

N-Ethyl-3-[BOC-L-(Tr-Gln)]-E-propenamide (0.040 g, 0.074 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.030 g, 0.073 mmol) usingthe procedure described in Example 28 for the preparation ofethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate to provideN-ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenamide (0.043 g, 70%) as awhite solid: mp=190° C. (dec); IR (thin film) 3283, 3067, 1693, 1642,1535 cm⁻¹; ¹H NMR (CDCl₃) δ 0.83 (d, 3H, J=9.0 Hz), 0.85 (d, 3H, J=9.0Hz), 1.14 (t, 2H, J=7.3 Hz), 1.21-1.32 (m, 1H), 1.37-1.52 (m, 2H),1.71-1.78 (m, 1H), 1.94-2.05 (m, 1H), 2.26 (t, 2H, J=7.3 Hz), 2.91 (dd,1H, J=13.8, 7.6 Hz), 3.16 (dd, 1H, J=13.8, 6.2 Hz), 3.26-3.35 (m, 2H),3.94-4.01 (m, 1H), 4.53-4.55 (m, 2H), 4.89-4.94 (m, 3H), 5.56-5.65 (m,2H), 6.51 (d, 1H, J=8.1 Hz), 6.60 (dd, 1H, J=15.1, 4.8 Hz), 6.81 (d, 1H,J=8.4 Hz), 7.02 (s, 1H), 7.10-7.36 (m, 26H); Anal. (C₅₁H₅₇N₅O₆) C, H, N.

Preparation of Products N-Ethyl-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenamide

Using the procedure described in Example 4 for the preparation ofcompound 3, N-ethyl-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenamide wasdeprotected to produce the product. mp=230° C. (dec), R_(f)=0.28 (10%MeOH in CH₂Cl₂); IR (KBr) 3404, 3075, 2943, 1692, 1643 cm⁻¹; ¹H NMR(DMSO-d₆) δ 0.78 (d, 3H, J=11.5 Hz), 0.80 (d, 3H, J=11.5 Hz), 1.02 (t,3H, J=7.3 Hz), 1.24-1.29 (m, 2H), 1.32-1.47 (m, 1H), 1.67-1.71 (m, 2H),2.03-2.08 (m, 2H), 2.77-2.85 (m, 1H), 2.99-3.16 (m, 3H), 3.91-3.98 (m,1H), 4.29-4.34 (m, 1H), 4.48-4.49 (m, 1H), 4.97 (d, 1H, J=12.5 Hz), 5.04(d, 1H, J=12.5 Hz), 5.85 (d, 1H, J=15.3 Hz), 6.43 (dd, 1H, J=15.4, 6.4Hz), 6.75 (s, 1H), 7.20 (bs, 7H), 7.30-7.34 (m, 4H), 7.41 (d, 1H, J=7.8Hz), 7.90 (d, 1H, J=7.8 Hz), 7.97 (t, 1H, J=5.1 Hz), 8.08 (d, 1H, J=8.1Hz); Anal. (C₃₂H₄₃N₅O₆) C, H, N.

Example 28 Preparation of Compound 8:Ethyl-2-Fluoro-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate Preparation ofIntermediate Ethyl-2-Fluoro-3-[BOC-L-(Tr-Gln)]-E-Propenoate

Sodium bis(trimethylsilyl)amide (0.264 mL of a 1.0 M solution in THF,0.264 mmol) was added to a solution oftriethyl-2-fluoro-2-phosphonoacetate (0.054 mL, 0.266 mmol) in THF (10mL) at −78° C., and the resulting solution was stirred for 15 minutes atthat temperature. BOC-L-(Tr-Glutaminal) (0.125 g, 0.264 mmol) in THF (10mL) was added via cannula, and the reaction mixture was stirred for 30minutes at −78° C. then was partitioned between 0.5 M HCl (100 mL) and a1:1 mixture of EtOAc and hexanes (2×100 mL). The organic layers weredried over Na₂SO₄ and were concentrated. Purification of the residue byflash column chromatography (30% EtOAc in hexanes) providedethyl-2-fluoro-3-[BOC-L-(Tr-Gln)]-E-propenoate (0.094 g, 63%) as a whitefoam: IR (thin film) 3324, 1724, 1670 cm⁻¹; ¹H NMR (CDCl₃) δ 1.33 (t,3H, J=7.2 Hz), 1.41 (s, 9H), 1.92-2.05 (m, 2H), 2.39 (t, 2H, J=7.2 Hz),4.28 (q, 2H, J=7.2 Hz), 5.00 (bs, 2H), 5.74 (dd, 1H, J=19.8, 8.6 Hz),6.78 (s, 1H), 7.14-7.32 (m, 15H); Anal. (C₃₃H₃₇FN₂O₅) C, H, N.

Preparation of IntermediateEthyl-2-Fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Propenoate

A solution of HCl in 1,4-dioxane (4 mL of a 4.0 M solution, 16 mmol) wasadded to a solution of ethyl-2-fluoro-3-[BOC-L-(Tr-Gln)]-E-propenoate(0.310 g, 0.553 mmol) in the same solvent (4 mL) at 23° C. The reactionmixture was stirred for 4 hours at 23° C., then was concentrated. Theresulting oil was dissolved in CH₂Cl₂, and CBZ-L-Leu-L-Phe (0.228 g,0.553 mmol), 1-hydroxybenzotriazole hydrate (0.112 g, 0.828 mmol),4-methylmorpholine (0.182 mL, 1.67 mmol), and1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (0.159 g,0.829 mmol) were added sequentially. The reaction mixture was stirredfor 12 hours at 23° C., then was partitioned between water (100 mL) andEtOAc (2×100 mL). The organic layers were dried over Na₂SO₄ and wereconcentrated. Purification of the residue by flash column chromatography(5% CH₃OH/CH₂Cl₂) affordedethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate (0.203 g,43%) as a white foam: IR (thin film) 3394, 3066, 1724, 1647 cm⁻¹; ¹H NMR(CDCl₃) δ 0.84 (d, 3H, J=5.9 Hz), 0.86 (d, 3H, J=6.2 Hz), 1.32 (t, 3H,J=7.0 Hz), 1.37-1.57 (m, 3H), 1.82-1.84 (m, 2H), 2.26-2.29 (m, 2H),2.97-2.99 (m, 2H), 3.99-4.05 (m, 1H), 4.26 (q, 2H, J=7.0 Hz), 4.46-4.49(m, 1H), 4.95 (s, 2H), 5.06 (d, 1H, J=6.5 Hz), 5.16-5.21 (m, 1H), 5.54(dd, 1H, J=19.9, 9.7 Hz), 6.55 (d, 1H, J=7.5 Hz), 6.79 (d, 1H, J=7.5Hz), 6.99 (s, 1H), 7.07-7.42 (m, 25H); Anal. (C₅₁H₅₅FN₄O₇) C, H, N.

Preparation of ProductEthyl-2-Fluoro-3-(CBZ-L-Leu-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 4 for the preparation ofcompound 3, ethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoatewas deprotected to produce the product. mp=210-211° C., R_(f)=0.57 (10%MeOH in CH₂Cl₂); IR (KBr) 3401, 3300, 3072, 2943, 1693, 1648 cm⁻¹; ¹HNMR (DMSO-d₆) δ 0.79 (d, 3H, J=10.9 Hz), 0.82 (d, 3H, J=10.9 Hz), 1.27(t, 3H, J=7.2 Hz), 1.32-1.49 (m, 3H), 1.65-1.80 (m, 2H), 1.99-2.06 (m,2H), 2.78-2.96 (m, 2H), 3.96-4.01 (m, 1H), 4.25 (q, 2H, J=7.2 Hz),4.39-4.41 (m, 1H), 4.97-5.07 (m, 3H), 5.65 (dd, 1H, J=21.2, 10.0 Hz),6.74 (s, 1H), 7.16-7.30 (m, 7H), 7.32-7.34 (m, 4H), 7.44 (d, 1H, J=8.1Hz), 7.94 (d, 1H, J=8.1 Hz), 8.03 (d, 1H, J=7.8 Hz). Anal. (C₃₂H₄₁FN₄O₇)C, H, N.

Example 29 Preparation of Compound 9:Methyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Sulfone Preparation ofIntermediate Methyl-(2-[BOC-L-(Tr-Gln)]-E-Vinyl)Sulfone

Sodium bis(trimethylsilyl)amide (1.04 mL of a 1.0 M solution in THF,1.04 mmol) was added to a solution of methanesulfonylmethyl-phosphinicacid diethyl ether (0.217 g, 0.943 mmol) in THF (30 mL) at −78° C., andthe resulting solution was stirred for 15 minutes at that temperature.BOC-L-(Tr-Glutaminal) (0.446 g, 0.944 mmol) in THF (15 mL) was added viacannula, and the reaction mixture was stirred for 30 minutes at −78° C.then was partitioned between 0.5 M HCl (100 mL) and a 1:1 mixture ofEtOAc and hexanes (2×100 mL). The organic layers were dried over Na₂SO₄and were concentrated. Purification of the residue by flash columnchromatography (40% hexanes in EtOAc) providedmethyl-(2-[BOC-L-(Tr-Gln)]-E-vinyl) sulfone (0.359 g, 69%) as a whitefoam: IR (thin film) 3348, 1688, 1495 cm⁻¹; ¹H NMR (CDCl₃) δ 1.43 (s,9H), 1.64-1.81 (m, 1H), 1.83-2.01 (m, 1H), 2.40 (t, 2H, J=6.7 Hz), 2.91(s, 3H), 4.35 (s, br, 1H), 5.01-5.04 (m, 1H), 6.42 (dd, 1H, J=15.0, 1.7Hz), 6.78 (s, 1H), 6.78 (dd, 1H, J=15.0, 5.0 Hz), 7.18-7.33 (m, 15H);Anal. (C₃₁H₃₆N₂O₅S) C, H, N.

Preparation of IntermediateMethyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Vinyl) Sulfone

Using the procedure described in Example 28 for the preparation ofethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,methyl-(2-[BOC-L-(Tr-Gln)]-E-vinyl) sulfone (0.359 g, 0.654 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.270 g, 0.655 mmol) toprovide methyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-vinyl) sulfone (0.160g, 29%) as a white foam: IR (thin film) 3296, 3061, 1649, 1529 cm⁻¹; ¹HNMR (CDCl₃) δ 0.84 (d, 3H, J=8.9 Hz), 0.86 (d, 3H, J=8.9 Hz), 1.24-1.36(m, 2H), 1.42-1.55 (m, 2H), 1.72-1.75 (m, 1H), 1.96-1.99 (m, 1H),2.23-2.32 (m, 2H), 2.85 (s, 3H), 2.97 (dd, 1H, J=13.8, 7.5 Hz), 3.13(dd, 1H, J=13.8, 6.1 Hz), 3.92-3.99 (m, 1H), 4.43-4.56 (m, 2H), 4.88 (s,br, 2H), 4.95 (d, 1H, J=5.9 Hz), 6.20 (d, 1H, J=14.9 Hz), 6.47 (d, 1H,J=7.2 Hz), 6.70 (dd, 1H, J=14.9, 4.4 Hz), 6.98 (d, 1H, J=8.1 Hz),7.09-7.38 (m, 25H).

Preparation of Product—Methyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Sulfone

Using the procedure described in Example 4 for the preparation ofcompound 3, methyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-vinyl)sulfone wasdeprotected to produce the product. mp=220° C. (dec), R_(f)=0.44 (10%MeOH in CH₂Cl₂); IR (KBr) 3413, 3284, 3049, 2951, 1690, 1649 cm⁻¹; ¹HNMR (DMSO-d₆) δ 0.79 (d, 3H, J=10.6 Hz), 0.81 (d, 3H, J=10.6 Hz),1.27-1.38 (m, 2H), 1.40-1.50 (m, 1H), 1.63-1.80 (m, 2H), 2.08 (t, 2H,J=7.5 Hz), 2.82-2.89 (m, 1H), 2.96 (s, 3H), 2.98-3.04 (m, 1H), 3.94-3.99(m, 1H), 4.45-4.53 (m, 2H), 4.98 (d, 1H, J=12.5 Hz), 5.05 (d, 1H, J=12.5Hz), 6.38 (d, 1H, J=14.9), 6.60 (dd, 1H, J=15.4, 5.1 Hz), 6.78 (s, 1H),7.17-7.31 (m, 7H), 7.34-7.36 (m, 4H), 7.43 (d, 1H, J=8.1 Hz), 8.01 (d,1H, J=8.1 Hz), 8.13 (d, 1H, J=8.1 Hz); Anal. (C₃₀H₄₀N₄O₇S) C, H, N.

Example 30 Preparation of Compound 10:Phenyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Sulfone Preparation ofIntermediate Phenyl-(2-[BOC-L-(Tr-Gln)]-E-Vinyl)Sulfone

Sodium bis(trimethylsilyl)amide (1.14 mL of a 1.0 M solution in THF,1.14 mmol) was added to a solution of benzenesulfonylmethyl-phosphinicacid diethyl ether (0.304 g, 1.04 mmol) in THF (20 mL) at −78° C., andthe resulting solution was stirred for 15 minutes at that temperature.BOC-L-(Tr-Glutaminal) (0.491 g, 1.04 mmol) in THF (10 mL) was added viacannula, and the reaction mixture was stirred for 30 minutes at −78° C.then was partitioned between 0.5 M HCl (100 mL) and a 1:1 mixture ofEtOAc and hexanes (2×100 mL). The organic layers were dried over Na₂SO₄and were concentrated. Purification of the residue by flash columnchromatography (gradient elution, 30-40% EtOAc in hexanes) providedphenyl-(2-[BOC-L-(Tr-Gln)]-E-vinyl)sulfone (0.540 g, 85%) as a whitefoam: IR (thin film) 3347, 2250, 1688, 1493 cm⁻¹; ¹H NMR (CDCl₃) δ 1.37(s, 9H), 1.73-1.81 (m, 1H), 1.83-1.94 (m, 1H), 2.38 (t, 2H, J=6.7 Hz),4.33 (s, br, 1H), 4.88-4.90 (m, 1H), 6.37 (dd, 1H, J=15.3, 1.6 Hz),6.79-6.86 (m, 2H), 7.17-7.32 (m, 15H), 7.49-7.54 (m, 2H), 7.58-7.63 (m,1H), 7.83-7.87 (m, 2H); Anal. (C₃₆H₃₈N₂O₅S) C, H, N.

Preparation of IntermediatePhenyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Vinyl) Sulfone

Using the procedure described in Example 28 for the preparation ofethyl-2-fluoro-3-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-propenoate,phenyl-(2-[BOC-L-(Tr-Gln)]-E-vinyl) sulfone (0.205 g, 0.336 mmol) wasdeprotected and coupled with CBZ-L-Leu-L-Phe (0.138 g, 0.335 mmol) toprovide phenyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-vinyl) sulfone (0.100g, 33%) as a white foam: IR (thin film) 3298, 3061, 1652, 1518 cm⁻¹; ¹HNMR (CDCl₃) δ 0.81 (d, 3H, J=6.9 Hz), 0.83 (d, 3H, J=6.9 Hz), 1.24-1.69(m, 5H), 1.91 (s, br, 1H), 2.16-2.31 (m, 2H), 2.91 (dd, 1H, J=13.5, 7.5Hz), 3.05 (dd, 1H, J=13.5, 6.7 Hz), 3.91-3.98 (m, 1H), 4.38-4.45 (m,1H), 4.54 (s, br, 1H), 4.87 (s, br, 1H), 5.06 (d, 1H, J=6.2 Hz), 6.12(d, 1H, J=15.3 Hz), 6.57 (d, 1H, J=7.2 Hz), 6.75 (dd, 1H, J=15.3, 4.4Hz), 6.85 (d, 1H, J=8.4 Hz), 7.05 (d, 1H, J=7.2 Hz), 7.10-7.37 (m, 24H),7.40-7.62 (m, 3H), 7.79-7.82 (m, 2H); Anal. (C₅₄H₅₆N₄O₇S) C, H, N.

Preparation of Product—Phenyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Sulfone

Using the procedure described in Example 4 for the preparation ofcompound 3, phenyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-vinyl) sulfone wasdeprotected to produce the product. mp=230° C. (dec), R_(f)=0.40 (10%MeOH in CH₂Cl₂); IR (KBr) 3400, 3288, 3062, 2960, 1685, 1644 cm⁻¹; ¹HNMR (DMSO-d₆) δ 0.78 (d, 3H, J=10.6 Hz), 0.81 (d, 3H, J=10.6 Hz),1.26-1.39 (m, 2H), 1.47-1.59 (m, 1H), 1.61-1.66 (m, 1H), 1.76-1.79 (m,1H), 2.04 (t, 2H, J=7.0 Hz), 2.77-2.96 (m, 2H), 3.95-4.00 (m, 1H),4.43-4.45 (m, 2H), 4.96 (d, 1H, J=12.6 Hz), 5.02 (d, 1H, J=12.6 Hz),6.33 (d, 1H, J=14.9 Hz), 6.74-6.81 (m, 2H), 7.11-7.18 (m, 7H), 7.20-7.38(m, 4H), 7.42 (d, 1H, J=7.8 Hz), 7.65 (d, 2H, J=7.8 Hz), 7.71 (d, 1H,J=7.5 Hz), 7.82 (d, 2H, J=6.9 Hz), 8.00 (d, 1H, J=7.8 Hz), 8.09 (d, 1H,J=8.1 Hz); Anal. (C₃₅H₄₂N₄O₇S) C, H, N.

Example 31 Preparation of Compound 11:Ethyl-2-Fluoro-3-[BOC-L-(Cyanomethyl)-Ala]-E-Propenoate Preparation ofIntermediate BOC-L-Gln-OMe

To a solution of BOC-L-Gln (20 g, 81 mmol) in 50 mL of EtOAC and MeOH at0° C. was added diazomethane in 250 mL of Et₂O with stirring. Theresulting yellow solution was stirred at 0° C. for 5 minutes and thenwarmed up to room temperature and stirred for 20 minutes. Argon gas wasthen bubbled through the yellow reaction mixture to remove excessdiazomethane. The crude product was concentrated and purified bycrystallization from methyl-tert-butyl ether. Yield 100%. ¹H NMR (CDCl₃)δ 1.45 (s, 9H), 1.96 (m, 1H), 2.21 (m, 1H), 2.36 (m, 2H), 3.76 (s, 3H),4.34 (m, 1H), 5.32 (m, 1H), 5.44 (bs, 1H), 6.16 (bs, 1H). Anal.(C₁₁H₂₀N₂O₅) C, H, N.

Preparation of Intermediate BOC-L-(Cyanomethyl)-Ala-OMe

To a solution of BOC-L-Gln-OMe (10 g, 38 mmol) in 100 mL of pyridine at0° C. was added 3.5 mL of POCl₃ dropwise. The reaction was warmed toroom temperature and stirred overnight. The reaction mixture was dilutedwith 100 mL EtOAc and washed with 1N HCl (2×50 mL). The organics werecombined and dried over Na₂SO₄, concentrated to yield the crude productwhich was purified by flash column chromatography (1:4 EtOAc/hexane) togive the product in 67% yield. ¹H NMR (CDCl₃) δ 1.45 (s, 9H), 2.03 (m,1H), 2.27 (m, 1H), 2.46 (m, 2H), 3.80 (s, 3H), 4.38 (m, 1H), 5.20 (m,1H).

Preparation of Intermediate BOC-L-(Cyanomethyl)-Alaninol

This compound was prepared in 84% yield from BOC-L-(cyanomethyl)-Ala-OMeusing the procedure described in Example 2 for the preparation ofCBZ-L-(N-Ac-amino)-alaninol. The compound was purified by flash columnchromatography (50:50 EtOAc/hexane). ¹H NMR (CDCl₃) δ 1.45 (s, 9H), 1.92(m, 2H), 2.19 (m, 1H), 2.46 (m, 2H), 3.71 (m, 3H), 4.83 (m, 1H). Anal.(C₁₀H₁₈N₂O₃0.4 H₂O) C, H, N.

Preparation of Intermediate BOC-L-(Cyanomethyl)-Alaninal

To a solution of oxalyl chloride (1.63 g, 12.57 mmol) in CH₂Cl₂ (30 mL)at −78° C. was added DMSO dropwise (2.01 g, 25.74 mmol). After theaddition, the reaction was stirred for 5 minutes. A solution ofBOC-L-(cyanomethyl)-alaninol (2.5 g, 11.7 mmol) in 20 mL was added at−78° C. with stirring. After 20 minutes, the reaction was treated withNEt₃ (8.15 mL, 58.5 mmol) and stirred for another 20 minutes. Water (40mL) was added at −60° C., and then the reaction was warmed up to roomtemperature. The water layer was separated and extracted with EtOAc(2×50 mL). The organic layers were combined and dried over MgSO₄, andthen concentrated to give 2.1 g crude product which was purified byflash column chromatography using a gradient of 3:7 EtOAc/hexane to 5:5EtOAc/hexane to give the aldehyde in 60% yield. ¹H NMR (CDCl₃) δ 1.37(m, 3H), 1.42 (s, 9H), 1.46 (s, 9H), 1.91 (m, 1H), 2.55-2.30 (m, 3H),4.25 (m, 1H), 5.27 (m, 1H), 9.63 (s, 1H).

Preparation of IntermediateEthyl-2-Fluoro-3-[BOC-L-(Cyanomethyl)-Ala]-E-Propenoate

A solution of triethyl 2-fluoro-phosphonoacetate (0.31 g, 1.27 mmol) in4 mL THF was cooled at −78° C. and then n-BuLi (0.56 mL of 2.5 Msolution in hexanes, 1.39 mmol) was added. The resulting solution wasstirred at −78° C. for 20 minutes, and then a solution ofBOC-L-(cyanomethyl)-alaninal (0.124 g, 0.58 mmol) in 2 mL THF was addedto the reaction mixture. The reaction was allowed to stir at −78° C. for1 hour and then warmed up to room temperature and stirred overnight.Aqueous 6 N HCl (10 mL) was added to the reaction, and the organic layerwas separated and washed with brine (2×10 mL) and concentrated. Thecrude product was purified by flash column chromatography (30:70EtOAc/hexane) to give 0.07 g. product (55% yield). ¹H NMR (CDCl₃) δ2.2-1.8 (m, 2H), 2.45 (m, 2H), 4.33 (m, 2H), 4.77 (m, 1H), 5.01 (m,1H,), 5.89 (m, 1H). Anal. (C₁₄H₂₁N₂O₄F0.15 H₂O) C, H, N. MS calcd forC₁₄H₂₁N₂O₄F (M+Na), found 323.

Production ofProduct—Ethyl-2-Fluoro-3-[CBZ-L-Leu-L-Phe-L-(Cyanomethyl)-Ala]-E-Propenoate

A solution of ethyl-2-fluoro-3-[BOC-L-(cyanomethyl)-Ala]-E-propenoate(0.055 g, 0.18 mmol) in 1 mL CH₂Cl₂ was cooled to 0° C., and 0.3 mL ofTFA was added. The reaction was then warmed to room temperature, stirredfor 3 hours, concentrated, and trace amounts of water were removed bytoluene azeotrope. This crude product was dissolved in 2 mL DMF and asolution of benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (BOP) (0.12 g, 0.27 mmol), CBZ-L-Leu-L-Phe (0.11 g,0.27 mmol), and Et₃N (0.078 mL, 0.54 mmol) was added at 0° C., and thereaction was stirred for 4 hours. This reaction was diluted withsaturated aqueous NaHCO₃ solution and extracted with EtOAc (3×15 mL).The organics layers were combined and dried with MgSO₄ and concentrated.The residue was purified by flash column chromatography using a solventgradient of 1% MeOH/CH₂Cl₂ to 5% MeOH/CH₂Cl₂ yielding the product in 37%(2-steps). Anal (C₃₂H₃₉N₄O₆F) C, H, N. HRMS calcd for C₃₂H₃₉N₄O₆F+Na617.2751 (M+Na), found 617.2738.

Example 32 Preparation of Compound 20:Diethyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl] Phosphonate Preparation ofIntermediate CBZ-L-(Tr-Gln)

CBZ-L-Gln (28.03 g, 100 mmol) was dissolved in 300 mL of glacial aceticacid. To this solution was added triphenylmethanol (26.83 g, 100 mmol),acetic anhydride (18.87 mL, 200 mmol), and 0.5 mL of sulfuric acid. Thereaction was heated to 55° C., stirring for one hour. After cooling toroom temperature the mixture was concentrated under reduced pressure toone-third the original volume. Ice water was added, and the productextracted with EtOAc. The organic layer was washed with water and brine,dried over MgSO₄, and concentrated. The crude product was recrystallizedfrom CH₂Cl₂/hexane, and the resulting crystals washed with Et₂O,yielding 37.27 g (71%) as a white solid: IR (KBr) 3418, 3295, 3059,3032, 2949, 2515, 1699, 1628, 1539, 1504, 1447, 1418, 1341, 1242, 1209,1061, 748 696 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.71 (m, 1H), 1.88 (m, 1H), 2.38(m, 2H), 3.97 (m, 1H) 5.04 (s, 2H), 7.14-7.35 (m, 20H), 7.52 (d, 1H,J=7.7 Hz), 8.60 (s, 1H).

Preparation of Intermediate CBZ-L-(Tr-Gln)OMe

CBZ-L-(Tr-Gln) (0.26 g, 0.5 mmol) was added to a stirring solution of0.25 mL of acetyl chloride in 5 mL of MeOH, and stirring was continuedat room temperature for 1 hour. The solvent was removed in vacuo, andthe residue dissolved in 100 ml CH₂Cl₂. The organic layer was washedwith water, saturated NaHCO₃, and brine followed by drying over Na₂SO₄.The crude product was purified on a short flash silica gel column,eluting with 20% EtOAc/hexane. The product (0.23 g, 84%) was obtained asa white solid: IR (KBr) 3405, 3277, 3057, 3034, 2953, 1724, 1643, 1532,1493, 1447, 1207, 1042, 750, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.16 (t, 1H,J=7.0 Hz), 1.77 (m, 1H), 1.97 (m, 1H), 3.61 (s, 3H), 4.99 (m, 1H), 5.03(s, 2H), 7.02-7.55 (m, 20H), 7.69 (d, 1H, J=7.7 Hz), 8.59 (s, 1H). Anal.(C₃₃H₃₂N₂O₅) C, H, N.

Preparation of Intermediate CBZ-L-(Tr-Glutaminol)

CBZ-L-(Tr-Gln)OMe (1.50 g, 2.79 mmol) was dissolved in 20 mL of THF and10 mL of EtOH. LiCl (0.24 g, 5.6 mmol) was added, and the mixturestirred for 10 minutes until all solids had dissolved. NaBH₄ (0.21 g,5.6 mmol) was added, and the reaction stirred overnight at roomtemperature. The solvents were removed in vacuo, the residue taken up inwater, and the pH was adjusted to 2-3 with 10% HCl. The product wasextracted with EtOAc, and the organic layer was washed with water andbrine before drying over MgSO₄. The crude product was purified on ashort flash silica gel column, eluting with an increasing gradient ofEtOAc/benzene, yielding 1.02 g (72%) of a white glassy solid: IR (KBr)3408, 3318, 3057, 3032, 2947, 1699, 1674, 1516, 1447, 1240, 1059, 752,698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.40 (m, 1H), 1.72 (m, 1H), 2.26 (m, 2H),3.17-3.50 (m, 3H), 4.64 (t, 1H, J=5.0 Hz), 5.00 (s, 2H), 7.00-7.40 (m,20H), 6.96 (d, 1H, J=8.5 Hz), 8.54 (s, 1H). Anal. (C₃₂H₃₂N₂O₄) C, H, N.

Preparation of Intermediate L-(Tr-Glutaminol)

This amino alcohol was prepared from CBZ--L-(Tr-glutaminol) in 98% yieldusing the procedure described in Example 2 for the preparation ofL-(N-Ac-amino)-alaninol. IR (KBR) 3255, 3057, 3016, 2916, 1642, 1527,1491, 1446, 1057, 1036, 750, 700, 636 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.29 (m,1H), 1.53 (m, 1H), 2.29 (m, 2H), 3.08 (m, 1H), 3.18 (m, 2H), 3.38 (bs,2H), 4.43 (bs, 1H), 7.14-7.28 (m, 15H), 8.62 (s, 1H). Anal. (C₂₄H₂₆H₂O₂)C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(Tr-Glutaminol)

Using the procedure described in Example 1 for the preparation ofCBZ-L-Leu-L-Phe-L-methioninol, this derivative was synthesized fromCBZ-L-Leu-L-Phe and L-Tr-glutaminol in 62% yield as a white solid: IR(KBr) 3302, 3057, 3032, 2951, 1954, 1885, 1657, 1520, 1238, 1045, 746,698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.79 (t, 6H, J=7.0 Hz), 1.30 (m, 2H), 1.44(m, 2H), 1.75 (m, 1H), 2.22 (m, 2H), 2.82 (m, 1H), 2.97 (m, 1H), 3.14(m, 1H), 3.25 (m, 1H), 3.63 (m, 1H), 3.95 (m, 1H), 4.48 (m, 1H), 4.65(t, 1H, J=5.0 Hz), 4.96 (d, 1H, J=13.0 Hz), 5.02 (d, 1H, J=13.0 Hz),7.07-7.33 (m, 25H), 7.42 (d, 1H, J=8.0 Hz), 7.66 (d, 1H, J=8.5 Hz), 7.86(d, 1H, J=8.0 Hz), 8.52 (s, 1H). Anal. (C₄₇H₅₂N₄O₅0.5 H₂O) C, H, N.

Preparation of Intermediate CBZ-L-Leu-L-Phe-L-(Tr-Glutaminal)

Using the procedure described in Example 1 for the preparation ofCBZ-L-Leu-L-Phe-L-methioninal, this aldehyde was synthesized fromCBZ-L-Leu-L-Phe-L-(Tr-glutaminol) in 92% yield as a white glassy solid,which was used immediately. ¹H NMR (DMSO-d₆) δ 0.79 (t, 6H, J=7.0 Hz),1.00-1.98 (m, 5H), 2.27 (m, 2H), 2.84 (m, 1H), 3.02 (m, 1H), 3.98 (m,2H), 4.58 (m, 1H), 4.99 (s, 2H), 7.14-7.32 (m, 25H), 7.39 (d, 1H, J=8.0Hz), 7.97 (d, 1H, J=8.5 Hz), 8.38 (d, 1H, J=8.0 Hz), 8.60 (s, 1H), 9.20(s, 1H).

Preparation of IntermediateDiethyl-(2-[CBZ-L-Leu-L-Phe-L-(Tr-Gln)]-E-Vinyl) Phosphonate

Tetraethyl methylenediphosphonate (0.21 mL, 0.86 mmol) was dissolved in10 mL of THF and cooled to 0° C. Potassium bis(trimethylsilyl)amide (0.5M in toluene) was added dropwise via syringe, and the reaction stirredat 0° C. for 30 minutes. After cooling the reaction to −30° C. asolution of CBZ-L-Leu-L-Phe-L-(Tr-glutaminol) (0.63 g, 0.82 mmol) in 6mL of THF was added dropwise. The reaction was allowed to warm slowly toroom temperature and stirred overnight. The solvent was removed byevaporation, and the crude product was purified by flash columnchromatography eluting with 1% (saturated anhydrous (NH₃/MeOH)/CHCl₃ toafford 0.50 g (68%) of a white crystalline solid: IR (KBr) 3289, 3059,3032, 2957, 1667, 1532, 1447, 1246, 1026, 968, 748, 698 cm⁻¹; ¹H NMR(DMSO-d₆) δ 0.78 (t, 6H, J=7.0 Hz), 1.20 (m, 6H), 1.15-1.78 (m, 5H),2.25 (m, 2H), 2.85 (m, 1H), 2.97 (m, 1H), 3.86-4.07 (m, 5H), 4.32 (m,1H), 4.51 (m, 1H), 4.95 (d, 1H, J=13.0 Hz), 5.02 (d, 1H, J=13.0 Hz),5.52 (t, 1H, J=19.0 Hz), 6.48 (t, 1H, J=19.0 Hz), 7.07-7.32 (m, 25H),7.41 (d, 1H, J=8.0 Hz), 7.97 (d, 1H, J=8.5 Hz), 8.05 (d, 1H, J=8.0 Hz),8.59 (s, 1H); MS (M+H) 901, (M−H) 899. Anal. (C₅₂H₆₁N₄O₈P2.5 H₂O) C, H,N.

Preparation of Product Diethyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Phosphonate

The protected amide diethyl-[2-(CBZ-L-Leu-L-Phe-L-Gln)-E-Vinyl]Phosphonate (0.469 g, 0.52 mmol) was dissolved in 10 mL of CH₂Cl₂.Triisopropylsilane (0.52 mL) was added as a triphenylmethyl cationscavenger. TFA (1.0 mL) was added, and the reaction was stirredovernight at room temperature. The reaction was poured into EtOAc andwashed with saturated NaHCO₃ solution. The organic layer was separatedand washed with water and brine followed by drying over MgSO₄. Theproduct was purified by flash column chromatography eluting with 2-3%MeOH/CHCl₃ to give in 67% yield of a white solid: IR (KBr) 3291, 3063,2955, 1647, 1541, 1236, 1026, 968, 746, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ0.79 (m, 6H), 1.21 (t, 6H, J=7.0 Hz), 1.28 (m, 2H), 1.52 (m, 1H), 1.63(m, 1H), 1.75 (m, 1H), 2.06 (m, 2H), 2.85 (m, 1H), 3.00 (m, 1H), 3.92(m, 5H), 4.34 (m, 1H), 4.50 (m, 1H), 4.97 (d, 1H, J=13.0 Hz), 5.04 (d,1H, J=13.0 Hz), 5.54 (t, 1H, J=19.0 Hz), 6.49 (t, 1H, J=19.0 Hz), 6.77(bs, 1H), 7.15-7.34 (m, 11H), 7.44 (d, 1H, J=8.0 Hz), 8.00 (d, 1H, J=8.5Hz), 8.03 (d, 1H, J=8.0 Hz); HRMS calcd for C₃₃H₄₈N₄O₈P 659.3210 (M+H),found 659.3223. Anal. (C₃₃H₄₈N₄O₈P) C, H, N.

Example 33 Preparation of Compound 29:Ethyl-3-[N-(1-Tr-4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-Gln]-E-PropenoatePreparation ofProduct—Ethyl-3-[N-(1-Tr-4-Methoxyindole-2-Carbonyl)-L-(4-Cl-Phe)-L-Gln]-E-Propenoate

This compound was prepared by the deprotection ofethyl-3-[N-(4-methoxyindole-2-carbonyl)-L-(4-Cl-Phe)-L-(Tr-Gln)]-E-propenoate,using the procedure described in Example 32 for the preparation ofcompound 20, but in the absence of triisopropylsilane. ¹H NMR (DMSO-d₆)δ 1.20 (t, 3H, J=7.0 Hz), 1.74 (m, 2H), 2.03 (t, 2H, J=8.0 Hz), 2.94 (m,2H), 3.89 (s, 3H), 4.11 (q, 2H, J=7.0 Hz), 4.46 (m, 1H), 4.60 (m, 1H),5.70 (d, 1H, J=15.0 Hz), 6.54 (d, 1H, J=7.8 Hz), 6.70 (dd, 1H, J=15.0,5.7 Hz), 6.75 (bs, 1H), 6.87 (d, 1H, J=8.5 Hz), 7.06 (m, 5H), 7.31 (m,18H), 7.72 (bs, 1H), 8.26 (d, 1H, J=8.2 Hz), 8.61 (d, 1H, J=8.1 Hz);HRMS calcd for C₄₇H₄₅N₄O₆Cl+Cs 929.2082 (M+Cs), found 929.2078 Anal.(C₄₇H₄₅N₄O₆C11.0 H₂O) C, H, N.

Example 34 Preparation of Compound 167:Ethyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-Gln]-E-PropenoatePreparation of Intermediate CBZ-L-Phe-L-(Tr-Glutaminol)

Using the procedure described in Example 16 for the preparation ofBOC-L-(4-Cl-Phe)-(Tr-glutaminol), CBZ-L-Phe-L-(Tr-glutaminol) wassynthesized from CBZ-L-Phe and L-(Tr-glutaminol) in 67% yield as a whiteglassy solid: IR (KBr) 3304, 3059, 3030, 2936, 1956, 1887, 1809, 1659,1495, 1446, 1246, 1036, 750, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.47 (m, 1H),1.72 (m, 1H), 2.26 (m, 2H), 2.75 (m, 1H), 2.94 (m, 1H), 3.18 (m, 1H),3.26 (m, 1H), 3.66 (m, 1H), 4.21 (m, 1H), 4.66 (m, 1H), 4.09 (m, 2H),7.15-7.30 (m, 25H), 7.43 (d, 1H, J=8.5 Hz), 7.72 (d, 1H, J=9.0 Hz), 8.49(s, 1H), Anal. (C₄₁H₄₁N₃O₅•1.0 H₂O) C, H, N.

Preparation of Intermediate L-Phe-L-(Tr-Glutaminol)

Using the procedure described in Example 2 for the preparation ofL-(N-Ac-amino)-alaninol, L-Phe-L-(Tr-Glutaminol) was synthesized fromCBZ-L-Phe-L-(Tr-glutaminol) in quantitative yield as a white glassysolid: IR (KBr) 3293, 3061, 3026, 2938, 2361, 1669, 1495, 1446, 752, 700cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.46 (m, 1H), 1.78 (m, 1H), 2.28 (m, 2H), 3.10(m, 2H), 3.21 (m, 1H), 3.25 (m, 1H), 3.62 (m, 1H), 3.86 (t, 1H, J=6.0Hz), 4.72 (m, 1H), 7.10-7.32 (m, 20H), 8.14 (d, 1H, J=8.0 Hz), 8.53 (s,1H). MS calcd for C₃₃H₃₅N₃O₃+H 522, found 522. Anal. (C₃₃H₃₅N₃O₃•0.55CH₂Cl₂) C, H, N.

Preparation of IntermediateBOC-L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-Glutaminol)

L-Phe-L-(Tr-Glutaminol) (0.65 g, 1.25 mmol) was dissolved in 5 mL ofDMF. Diisopropylethylamine (0.44 mL, 2.5 mmol) was added, followed by0.29 g (1.25 mmol) of BOC-L-α-t-butylglycine. The reaction was cooled to0° C. and HATU [O-(7-azabenztriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate] (0.48 g, 1.25 mmol) was added. The reaction mixturewas allowed to warm to rt at which time the DMF was removed in vacuo.The residue was dissolved with EtOAc, and the organic phase washedconsecutively with 10% aq HCl solution, sat. NaHCO₃ solution, H₂O, andbrine. The solvent was dried (MgSO₄) and filtered, and the residuepurified by flash silica gel chromatography using a gradient solventsystem (0-1.5% MeOH/CHCl₃) to give 0.78 g (85%) of a white amorphoussolid: IR (KBr) 3314, 2967, 1657, 1495, 1368, 1246, 1169, 1057, 752, 700cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.78 (s, 9H), 1.37 (s, 10H), 1.72 (m, 1H), 2.23(m, 2H), 2.80 (m, 1H), 2.92 (m, 1H), 3.08 (m, 1H), 3.21 (m, 1H), 3.60(m, 1H), 3.83 (d, 1H, J=9.0 Hz), 4.55 (m, 1H), 4.59 (t, 1H, J=5.5 Hz),6.42 (d, 1H, J=9.0 Hz), 7.14-7.28 (m, 20H), 7.67 (d, 1H, J=8.0 Hz), 7.95(d, 1H, J=8.0 Hz), 8.45 (s, 1H); Anal. (C₄₄H₅₄N₄O₆•1.0 H₂O) C, H, N.

Preparation of Intermediate L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-Glutaminol)Hydrochloride Salt

BOC-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminol) (0.745 g, 1.01 mmol) wasdissolved in 2 mL of CH₂Cl₂ followed by 20 mL of Et₂O. Dry HCl gas wascarefully bubbled into the solution until the white solid stoppedprecipitating. The reaction mixture was concentrated, and 2-3 mL of THFwas added which redissolved the white solids. Thin layer chromatographyindicated that the reaction went to completion. The THF was removedunder vacuum and white solids were washed thoroughly with an excess ofEt₂O and dried to yield L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminol)hydrochloride salt in 95% yield. IR(KBr) 3258, 3057, 2967, 1661, 1520,700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.95 (s, 9H), 1.44 (m, 1H), 1.72 (m, 1H),2.13 (m, 1H), 2.25 (m, 1H), 2.97 (m, 2H), 3.06 (m, 1H), 3.15 (m, 1H),3.60 (m, 2H), 4.25 (bs, 1H), 4.55 (m, 1H), 7.13-7.27 (m, 20H), 7.89 (d,1H, J=8.0 Hz), 8.13 (bs, 2H), 8.49 (s, 1H), 8.61 (d, 1H, J=7.7 Hz);Anal. (C₃₉H₄₆H₄O₄•HCl·1.0 H₂O) C, H, N.

Preparation of IntermediateEthylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-Glutaminol)

L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-glutaminol) hydrochloride salt (0.61 g,0.91 mmol) was dissolved in 9 mL of CH₂Cl₂. Triethylamine (0.26 mL, 1.87mmol) was added, followed by the addition of 0.097 g (0.91 mL) of ethylchlorothiolformate. After stirring for five minutes at rt, the solventwas removed under reduced pressure, and the residue was purified bycolumn chromatography on silica gel eluting with a gradient solventsystem (0-2% MeOH/CHCl₃) to give 0.47 g (71%) of a white amorphoussolid: IR(KBr) 3300, 3059, 3026, 2967, 1649, 1493, 1194, 750, 698 cm⁻¹;¹H NMR (DMSO-d₆) 0.83 δ (s, 9H), 1.16 (t, 3H, J=7.0 Hz), 1.42 (m, 1H),1.69 (m, 1H), 2.23 (m, 2H), 2.75 (q, 2H, J=7.0 Hz), 2.80 (m, 1H), 2.96(m, 1H), 3.08 (m, 1H), 3.18 (m, 1H), 3.62 (m, 1H), 4.25 (d, 1H, J=9.0Hz), 4.48 (m, 1H), 5.75 (t, 1H, J=5.0 Hz), 7.10-7.28 (m, 20H), 7.60 (d,1H, J=8.5 Hz), 7.93 (d, 1H, J=9.0 Hz), 8.09 (d, 1H, J=7.7 Hz), 8.48 (s,1H); Anal. (C₄₂H₅₀N₄O₅S) C, H, N.

Preparation of IntermediateEthylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-Glutaminal)

Using the general procedure described in Example 1 for the preparationof CBZ-L-Leu-L-Phe-L-methioninal (sulfoxide),ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminal) wassynthesized fromethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminol) inquantitative yield and isolated as a white amorphous solid and usedwithout further purification: ¹H NMR (DMSO-d₆) δ 0.83 (s, 9H), 1.16 (t,3H, J=7.0 Hz), 1.55 (m, 1H), 1.86 (m, 1H), 2.26 (m, 2H), 2.74 (q, 2H,J=76.0 Hz), 2.85 (m, 1H), 2.98 (m, 1H), 3.90 (m, 1H), 4.25 (d, 1H, J=9.0Hz), 4.59 (m, 1H), 7.14-7.28 (m, 20H), 7.93 (d, 1H, J=9.0 Hz), 8.18 (d,1H, J=7.7 Hz), 8.38 (d, 1H, J=6.6 Hz), 8.52 (s, 1H), 9.13 (s, 1H).

Preparation of IntermediateEthyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide]-E-propenoate,ethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-Gln)]-E-propenoatewas synthesized fromethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminal) (0.22 g,0.30 mmol) to give 0.28 g of material contaminated withtriphenylphosphone oxide which was used without further purification:white amorphous solid: 1H NMR (DMSO-d₆) δ 0.83 (s, 9H), 1.21 (m, 6H),1.60 (m, 2H), 2.25 (m, 2H), 2.74 (q, 2H, J=7.0 Hz), 2.82 (m, 1H), 2.92(m, 1H), 4.09 (q, 2H, J=7.0 Hz), 4.25 (d, 1H, J=9.0 Hz), 4.34 (m, 1H),4.52 (m, 1H), 5.53 (d, 1H, J=15.5 Hz), 6.63 (dd, 1H, J=15.5, 5.5 Hz),7.08-7.28 (m, 20H), 7.93 (d, 1H, J=9.0 Hz), 8.07 (d, 1H, J=7.7 Hz), 8.16(d, 1H, J=7.7 Hz), 8.51 (s, 1H).

Preparation ofProduct—Ethyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-Gln]-E-Propenoate

Ethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-Gln)]-E-propenoate,impure with triphenylphosphine oxide (0.28 g), was dissolved in 6mL ofCH₂Cl₂. TFA (0.6 mL) was added, and the reaction stirred at rt for 4hours. The reaction was poured into an EtOAc/sat. NaHCO₃ solution andagitated until white solids began to precipitate out of the organiclayer. The aqueous layer was separated, and the solids were filtered andwashed with EtOAc to give 0.074 g of a white solid (45% yield from theethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminal); 2 steps):IR(KBr) 3302, 2967, 1645, 1541, 1196 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.83 (s,9H), 1.18 (m, 6H), 1.67 (m, 2H), 2.03 (m, 2H), 2.75 (q, 2H, J=7.0 Hz),2.86 (m, 1H), 2.93 (m, 1H), 4.10 (q, 2H, J=7.0 Hz), 4.25 (d, 1H, J=9.0Hz), 4.35 (m, 1H), 4.49 (m, 1H), 5.55 (d, 1H, J=15.5 Hz), 6.64 (dd, 1H,J=15.5, 5.5 Hz), 6.73 (bs), 7.19 (m, 6H), 7.97 (d, 1H, J=8.5 Hz), 8.07(d, 1H, J=8.0 Hz), 8.15 (d, 1H, J=7.7 Hz); HRMS calcd for C₂₇H₄₀N₄O₆S+Cs681.1723, found 681.1738. Anal. (C₂₇H₄₀N₄O₆S) C, H, N.

Example 35 Preparation of Compound 168;Ethyl-2-Methyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-Gln]-E-PropenoatePreparation of IntermediateEthyl-2-Methyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Met (sulfoxide)]-E-propenoate,ethyl-2-methyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-Cln)]-E-propenoatewas synthesized fromethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminal) (0.22 g,0.30 mmol) and (carbethoxyethylidene)triphenylphosphorane (0.14 g, 0.37mmol). The product (0.31 g), a white amorphous solid, contaminated withtriphenylphosphine oxide, was isolated after column chromatography andused without further purification: ¹H NMR (DMSO-d₆) δ 0.83 (s, 9H), 1.18(m, 6H), 1.54 (m, 1H), 1.66 (m, 1H), 1.73 (s, 3H), 2.21 (m, 2H), 2.75(q, 2H, J=7.0 Hz), 2.80 (m, 1H), 2.88 (m, 1H), 4.12 (q, 2H, J=7.0 Hz),4.24 (d, 1H, J=9.0 Hz), 4.44 (m, 2H), 6.27 (d, 1H, J=8.5 Hz), 7.13-7.27(m, 20H), 7.95 (d, 1H, J=9.0 Hz), 8.03 (d, 1H, J=8.0 Hz), 8.09 (d, 1H,J=7.0 Hz), 8.51 (s, 1H).

Preparation ofProduct—Ethyl-2-Methyl-3-[Ethylthiocarbonyl-L-α-(t-Butyl-Gly)-L-Phe-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the preparation ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-2-methyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoatewas synthesized fromethyl-2-methyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-Gln)]-E-propenoateand isolated as a white glassy solid after purification by columnchromatography on silica gel using a gradient solvent system (0-2%MeOH/CHCl₃) (58% yield; two steps fromethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-(Tr-glutaminal): IR (KBr)3302, 2967, 1647, 1541, 1261, 1202 cm⁻¹; ¹H NMR (DMSO-d₆) δ 0.83 (s,9H), 1.18 (m, 6H), 1.65 (m, 1H), 1.69 (m, 1H), 1.77 (s, 3H), 2.00 (m,2H), 2.75 (q, 2H, J=7.0 Hz), 2.86 (m, 2H), 4.12 (q, 2H, J=7.0 Hz), 4.24(d, 1H, J=9.0 Hz), 4.42 (m, 2H), 6.26 (d, 1H, J=8.5 Hz), 6.71 (bs, 1H),7.15 (m, 6H), 7.96 (d, 1H, J=9.0 Hz), 8.03 (d, 1H, J=7.7 Hz), 8.07 (d,1H, J=7.0 Hz); HRMS calcd for C₂₈H₄₂N₄O₆S+Cs 695.1879, found 695.1864.Anal. (C₂₈H₄₂N₄O₆S•0.2 CHCl₃) C, H, N.

Example 36 Preparation of Compound 178:Ethyl-3-[Cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-Gln]-E-PropenoatePreparation of Intermediate BOC-L-(S-Me-Pen)-L-Phe-L-(Tr-Glutaminol)

L-Phe-L-(Tr-Glutaminol) (0.64 g, 1.25 mmol) was dissolved in 4 mL ofDMF. Diisopropylethylamine (0.43 mL, 2.46 mmol) was added, followed byBOC-S-methyl-L-penicillamine (0.32 g, 1.25 mmol; generated from theBOC-S-methyl-L-penicillamine dicyclohexylammonium salt (Sigma Chemical,St. Louis, Mo.) and aq HCl/EtOEt extraction and drying by benzeneazeotrope). The solution was cooled to 0° C., HATU(O-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate] (0.468 g, 1.25 mmol) was added, and the reactionmixture was allowed to warm to rt. The DMF was then removed in vacuo,the residue was dissolved with EtOAc, and the organic phase was washedconsecutively with 10% HCl solution, sat NaHCO₃, H₂O, and brine. Theorganic phase was dried over MgSO₄, filtered, and concentrated to give aresidue which was purified by column chromatography on silica gel usinga gradient solvent system (0-1% MeOH/CHCl₃) to yield 0.76 g (81%) of awhite amorphous solid: IR (KBr) 3308, 2937, 1695, 1677, 1506, 1493,1448, 1367, 1246, 1165, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.07 (s, 3H), 1.19(s, 3H), 1.37 (s, 9H), 1.66-1.75 (m, 2H), 1.94 (s, 3H), 2.19-2.25 (m,2H), 2.78-2.83 (m, 1H), 2.95-3.01 (m, 1H), 3.06-3.12 (m, 1H), 3.19-3.23(m, 1H), 3.62-3.65 (m, 1H), 4.12 (d, 1H, J=3.0 Hz), 4.48-4.55 (m, 1H),4.59-4.62 (m, 1H), 6.50 (d, 1H, J=9.0 Hz), 7.14-7.28 (m, 20H), 7.62 (d,1H, J=6.0 Hz), 8.21 (d, 1H, J=6.0 Hz), 8.47 (s, 1H). MS calcd forC₄₄H₅₄N₄O₆S+H 767, found 767.

Preparation of Intermediate L-(S-Me-Pen)-L-Phe-L-(Tr-Glutaminol)Hydrochloride Salt

To a solution of BOC-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol) (0.69 g, 0.91mmol) in 6 mL of 1,4-dioxane was added 4 mL of 4M HCl/1,4-dioxane. Thereaction mixture was stirred at rt for 3 h under an argon atmosphere. Atthis time the solvent was removed in vacuo to give 0.61 g (97%) of awhite solid which was used without further purification: IR (KBr) 3313,3057, 2926, 1664, 1493, 1448, 750, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.18 (s,3H), 1.39 (s, 3H), 1.66-1.78 (m, 2H), 2.01 (s, 3H), 2.06-2.15 (m, 1H),2.27-2.39 (m, 1H), 2.83-3.08 (m, 2H), 3.14-3.29 (m, 2H), 3.33-3.40 (m,3H), 3.59-3.68 (m, 1H), 3.84-3.89 (m, 1H), 7.13-7.27 (m, 20H), 7.91 (d,1H, J=9.0 Hz), 8.15-8.26 (m, 2H), 8.52 (s, 1H), 8.76 (d, 1H, J=6.0 Hz).

Preparation of IntermediateCyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-Glutaminol)

A solution of cyclopentyl chlorothiolformate (0.133 g, 0.81 mmol),prepared as described in Example 37, in 2 mL of CH₂Cl₂ was addeddropwise to a solution of L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol)hydrochloride salt (0.57 g, 0.81 mmol) in 10 mL of CH₂Cl₂. To thissolution was added 0.24 mL (1.7 mmol) of Et₃N. The reaction mixture wasstirred for 15 min at rt, and the solvent was removed under vacuum. Theresidue was purified by column chromatography on silica gelchromatography using a gradient solvent system (0-2% MeOH/CHCl₃) to give0.512 g (80%) of a white amorphous solid: IR (KBr) 3358, 2939, 1649,1516, 1448, 1190, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.13 (s, 3H), 1.23 (s,3H), 1.37-1.63 (m, 10H), 1.96 (s, 3H), 1.98-2.01 (m, 1H), 2.16-2.33 (m,1H), 2.7-2.89 (m, 1H), 3.07-3.23 (m, 2H), 3.24-3.28 (m, 1H), 3.53-3.57(m, 1H), 3.59-3.66 (m, 1H), 4.37-4.47 (m, 1H), 4.54-4.60 (m, 2H),7.14-7.28 (m, 20H), 7.55 (d, 1H, J=9.0 Hz), 7.99 (d, 1H, J=9.0 Hz), 8.36(d, 1H, J=6.0 Hz), 8.49 (s, 1H). MS calcd for C₄₅H₅₄N₄O₅S₂+H 795, found795.

Preparation of IntermediatesCyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-Glutaminal) &Cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-(Tr-Glutaminal)

Cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol) (0.46 g,0.58 mmol) was dissolved in 10 mL of any DMSO. o-Iodoxybenzoic acid(0.48 g, 1.73 mmol) was added, and the reaction mixture was stirred atrt for 3 h. The DMSO was removed under high vacuum. The residue wastwice diluted with CH₂Cl₂ and the solvent was evaporated to remove anyresidual DMSO. The residue was diluted with EtOAc, and triturated toform a white solid which was filtered off. The filtrate was washed withan aq 10% Na₂S₂O₃/10% NaHCO₃ solution, water and brine and dried overMgSO₄. Filtration and concentration gave 0.40 g (87%) of a white glassysolid which was used without further purification. The product was shownto be a mixture of the sulfide and sulfoxide by NMR analysis. ¹H NMR(DMSO-d₆) (mixture of sulfide and sulfoxide) δ 1.12 (s), 1.24 (s), 1.32(s), 1.45-1.66 (m), 1.95-2.13 (m), 2.29 (s), 2.40 (s), 2.53 (s),2.82-2.87 (m), 2.99-3.23 (m). 3.52-3.57 (m), 3.95-4.03 (m), 4.55-4.83(m), 7.14-7.28 (m), 7.89-8.06 (m), 8.41-8.58 (m), 9.15 (s), 9.18 (s).

Preparation of IntermediatesEthyl-3-[Cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-Gln)]-E-Propenoate&Ethyl-3-(Cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-[Tr-Gln])-E-Propenoate

The mixture ofcyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminal) andcyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-(Tr-glutaminal) (0.40 g,approximately 0.51 mmol) was dissolved in 10 mL of anh THF. To thissolution was added (carbethoxymethylene) triphenylphosphorane (0.21 g,0.61 mmol), and the reaction mixture was stirred overnight at rt. Thesolvent was removed in vacuo, and the residue was purified by columnchromatography on silica gel using a gradient solvent system (0-2%MeOH/CHCl₃) to give 0.184 g of the sulfide product and 0.132 g sulfoxideproduct (contaminated with triphenylphosphine oxide):Ethyl-3-[cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-Gln)]-E-propenoate:¹H NMR (DMSO-d₆) δ 1.14 (s, 3H), 1.21 (t, 3H, J=6.0 Hz), 1.24 (s, 3H),1.46-1.68 (m, 10H), 1.96 (s, 3H), 2.25-2.31 (m, 2H), 2.78-2.85 (m, 1H),2.96-3.00 (m, 1H), 3.54-3.72 (m, 1H), 4.05-4.13 (m, 2H), 4.32-4.47 (m,1H), 4.49-4.55 (m, 1H), 4.56-4.59 (m, 1H), 5.57 (d, 1H, J=15.0 Hz), 6.64(dd, 1H, J=15.0, 3.0 Hz), 7.13-7.26 (m, 20H), 7.99-8.04 (m, 2H), 8.45(d, 1H, J=9.0 Hz), 8.55 (s, 1H).Ethyl-3-(cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-[Tr-Gln])-E-propenoate:¹H NMR (DMSO-d₆) (mixture of diastereomers): δ 1.11-1.15 (m), 1.19-1.23(m), 1.35-1.66 (m), 1.98-2.00 (m), 2.18-2.35 (m), 2.41 (s), 2.64-2.83(m), 2.89-3.02 (m), 3.51-3.56 (m), 4.11 (q, J=6.0 Hz), 4.34-4.40 (m),4.48-4.59 (m), 4.63-4.66 (m), 5.51-5.57 (m), 6.61-6.68 (m), 7.13-7.28(m), 8.12-8.24 (m), 8.42-8.53 (m), 8.55-8.57 (m).

Preparation ofProduct—Ethyl-3-[Cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-Gln]-E-Propenoate

Ethyl-3-[cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-Gln)]-E-propenoate(0.184 g) was dissolved in 10 mL CH₂Cl₂. To this solution was added 1 mLof trifluoroacetic acid, and the reaction mixture was stirred at rtovernight. The solvent was removed under vacuum and the residue waspurified by column chromatography on silica gel using a gradient solventsystem (0-2% MeOH/CHCl₃) to give 0.044 g (24%; 3 steps fromcyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol)) as a whiteamorphous solid: IR (KBr) 3296, 2984, 1787, 1655, 1560, 1541, 1280, 1194cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.14 (s, 3H), 1.21 (t, 3H, J=6.0 Hz), 1.25 (s,3H), 1.40-1.70 (m, 10H), 2.02 (s, 3H), 2.05-2.24 (m, 2H), 2.79-2.86 (m,1H), 2.93-3.00 (m, 1H), 3.43-3.55 (m, 1H), 4.09 (q, 2H, J=6.0 Hz),4.31-4.36 (m, 1H), 4.43-4.50 (m, 1H), 4.56 (d, 1H, J=6.0 Hz), 5.58 (d,1H, J=15.0 Hz), 6.65 (dd, 1H, J=15.0, 6.0 Hz), 6.75 (bs, 1H), 7.15-7.21(m, 6H), 7.99-8.06 (m, 2H), 8.45 (d, 1H, J=6.0 Hz). HRMS calcd forC₃₀H₄₄N₄O₆S₂+Cs 753.1757, found 753.1737. Anal. (C₃₀H₄₄N₄O₆S₂) C, H, N,S.

Example 37 Preparation of Compound 173:Ethyl-3-[Cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-Gln]-E-PropenoatePreparation of Intermediate Cyclopentyl Chlorothiolformate

Cyclopentanethiol (10.7 mL, 0.1 mol) was dissolved in 200 mL of CH₂Cl₂.Triphosgene (11.13 g, 37.5 mmol) was added and the reaction mixture wascooled to 0° C. Et₃N (14.1 mL, 0.1 mol) was added dropwise, and thereaction was allowed to warm to room temperature over a period of onehour. The solvent was carefully removed under reduced pressure at 20° C.due to the volatility of the product. The resulting residue was taken upin Et₂O, and the solids were filtered and washed with more Et₂O. Thesolvent was again carefully removed under reduced pressure, and the wasproduct purified by distillation (85% yield): colorless liquid (bp70-74° C.; 1 torr): IR(neat) 1756, 830 cm⁻¹; ¹H NMR (benzene-d₆) δ1.01-1.23 (m, 6H), 1.49-1.60 (m, 2H), 3.20-3.29 (m, 1H).

Preparation of Intermediate BOC-L-(S-Ph-Cys)

To a suspension of 19.73 g (0.1 mol) L-(S-Ph-Cys) (purchased from DavosChemical Corp., Englewood Cliffs, N.J.) in 72 mL of tert-butanol wasadded a solution of NaOH (4.1 g, 0.1025 mol) in 100 mL H₂O. Once thesuspension became a clear solution di-tert-butyl dicarbonate (22.92 g,0.105 mol) was added. The clear solution became a slurry and was allowedto stir at rt overnight. At this time the turbid solution was washedtwice with pet. ether. The organic layer was washed 3 times with a satNaHCO₃ solution and the aqueous layers were combined. The aqueous layerwas then carefully acidified to pH 2-3 with a sat KHSO₄ solution andextracted with a large excess of Et₂O. The organic phase was dried overNa₂SO₄, filtered and concentrated under vacuum to give 27.4 g (92%) ofBOC-L-(S-Ph-Cys) as white solid. Any residual H₂O and/or tert-butanolwas removed by benzene azetrope before using the material. ¹H NMR(DMSO-d₆) δ 1.36 (s, 9H), 3.10 (dd, 1H, J=13.6, 9.6 Hz), 3.34 (dd, 1H,J=13.6, 4.4 Hz), 4.01 (m, 1H), 7.20 (m, 2H), 7.34 (m, 3H), 12.82 (bs,1H).

Preparation of Intermediate BOC-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol)

BOC-L-(S-Ph-Cys) (0.45 g, 1.5 mmol) was dissolved in 2 mL of DMF and 2mL of CH₂Cl₂. To this solution was added N-hydroxysuccinimide (0.17 g,1.5 mmol), followed by dicyclohexylcarbodiimide (0.31 g, 1.5 mmol). Thereaction was stirred at rt for 2 h. The mixture was then filtered into aseparate flask containing L-Phe-L-(Tr-glutaminol) (0.78 g, 1.5 mmol)dissolved in 4 mL of DMF and 2 mL of CH₂Cl₂. The reaction mixture wasstirred overnight and the solvent was removed in vacuo. The residue waspurified by column chromatography on silica gel using a gradient solventsystem (0-2% MeOH/CHCl₃) to give 1.06 g (88%) of a white amorphoussolid: IR (KBr) 3304, 3061, 2972, 2928, 1645, 1516, 1493, 1367, 1248,1165, 1024, 742, 698 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.35 (s, 9H), 1.35-1.43(m, 1H), 1.70-1.74 (m, 1H), 2.20-2.33 (m, 2H), 2.82-2.92 (m, 1H),2.93-3.10 (m, 1H), 3.11-3.23 (m, 2H), 3.25-3.32 (m, 2H), 3.58-3.68 (m,1H), 3.80-3.98 (m, 1H), 4.58-4.64 (m, 1H), 4.65-4.77 (m, 1H), 7.14-7.30(m, 26H), 7.55 (d, 1H, J=6.0 Hz), 7.83 (d, 1H, J=6.0 Hz), 8.51 (s, 1H),MS calcd for C₄₇H₅₂N₄O₆S+H 801, found 801.

Preparation of Intermediate L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol)Hydrochloride Salt

Using the procedure described in Example 36 for the preparation ofL-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol) hydrochloride salt,L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol) hydrochloride salt was synthesizedfrom BOC-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol) to give 0.182 g of whitesolid which was used without further purification: IR (KBr) 3325, 3057,2949, 1685, 1655, 1560, 1493, 1448, 746, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ1.41-1.67 (m, 1H), 1.69-1.81 (m, 1H), 2.26-2.44 (m, 2H), 2.86-2.97 (m,1H), 2.98-3.23 (m, 1H), 3.25-3.43 (m, 4H), 3.60-3.84 (m, 2H), 4.02-4.20(m, 1H), 4.44-4.60 (m, 1H), 7.08-7.48 (m, 25H), 7.87 (d, 1H, J=6.0 Hz),8.46 (br, 3H), 8.55 (s, 1H), 8.87 (d, 1H, J=6.0 Hz).

Preparation of IntermediateCyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-Glutaminol)

Using the procedure described in Example 36 for the preparation ofcyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol),cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol) wassynthesized from L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminol) hydrochloride saltin 75% yield: white amorphous solid: IR (KBr) 3288, 3059, 2960, 1637,1494, 1448, 1205, 746, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.42-1.98 (m, 10H),1.99-2.26 (m, 1H), 2.48-2.50 (m, 1H), 2.96-2.98 (m, 1H), 3.01-3.19 (m,1H), 3.19-3.55 (m, 6H), 3.64-3.85 (m, 1H), 4.36-4.40 (m, 1H), 4.46-4.58(m, 1H), 7.14-7.30 (m, 25H), 7.68 (d, 1H, J=6.0 Hz), 8.01 (d, 1H, J=6.0Hz), 8.41 (d, 1H, J=6.0 Hz), 8.52 (s, 1H). MS calcd for C₄₈H₅₂N₄O₅S₂+H829, found 829.

Preparation of IntermediateCyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-Glutaminal)

Using the procedure described in Example 36 for the preparation ofcyclopenthylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminal) andcyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-(Tr-glutaminal),cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminal) wassynthesized fromcyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L(Tr-glutaminol) in 98%yield: white amorphous solid used without further purification: ¹H NMR(DMSO-d₆) δ 1.45-1.70 (m, 8H), 2.02-2.28 (m, 3H), 2.35-2.51 (m, 1H),2.95-3.02 (m, 2H), 3.04-3.22 (m, 1H), 3.24-3.36 (m, 1H), 3.56-3.59 (m,1H), 4.02-4.08 (m, 1H), 4.47-4.59 (m, 1H), 4.60-4.80 (m, 1H), 7.20-7.36(m, 25H), 8.22 (d, 1H, J=6.0 Hz), 8.43-8.48 (m, 2H), 8.65 (s, 1H), 9.27(s, 1H).

Preparation of IntermediateEthyl-3-[Cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide)]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-Gln)]-E-propenoatewas synthesized fromcyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminal) to give0.26 g of material contaminated with triphenylphosphine oxide (aftercolumn chromatography) which was used without further purification: ¹HNMR (DMSO-d₆) δ 1.19 (t, 3H, J=6.0 Hz), 1.47-1.59 (m, 10H), 1.93-2.23(m, 1H), 2.25-2.34 (m, 1H), 2.83-2.93 (m, 1H), 2.95-3.16 (m, 1H),3.19-3.29 (m, 2H), 3.51-3.56 (m, 1H), 4.09 (q, 2H, J=6.0 Hz), 4.35-4.44(m, 2H), 4.46-4.48 (m, 1H), 5.64 (d, 1H, J=15.0 Hz), 6.68 (dd, 1H,J=15.0, 3.0 Hz), 7.13-7.29 (m, 25H), 8.07 (d, 1H, J=6.0 Hz), 8.13 (d,1H, J=6.0 Hz), 8.42 (d, 1H, J=6.0 Hz), 8.58 (s, 1H).

Preparation ofProduct-Ethyl-3-[Cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the prepartion ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-Gln]-E-propenoatewas synthesized fromehtyl-3-[cyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-Gln)]-E-propenoatein 35% yield (2 steps fromcyclopentylthiocarbonyl-L-(S-Ph-Cys)-L-Phe-L-(Tr-glutaminal)): whiteamorphous solid: IR (KBr) 3294, 1712, 1655, 1633, 1545, 1203, 738, 700cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.19 (t, 3H, J=6.0 Hz), 1.56-1.76 (m, 10H),1.98-2.08 (m, 2H), 2.84-2.99 (m, 2H), 3.17-3.39 (m, 2H), 3.51-3.76 (m,1H), 4.08 (q, 2H, J=6.0 Hz), 4.39-4.45 (m, 3H), 5.64 (d, 1H, J=15.0 Hz),6.69 (dd, 1H, J=15.0, 3.0 Hz), 6.77 (bs, 1H), 7.18-7.32 (m, 11H), 8.08(d, 1H, J=6.0 Hz), 8.18 (d, 1H, J=6.0 Hz), 8.43 (d, 1H, J=6.0 Hz), HRMScalcd for C₃₃H₄₂N₄O₆S₂+Cs 787.1600, found 787.1618. Anal. (C₃₃H₄₂N₄O₆S₂)C, H, N, S.

Example 38 Preparation of Compound 174:Ethyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-PropenoatePreparation of Intermediate Fmoc-L-(4-Me-Phe)-L-(Tr-Glutaminol)

Using the procedure described in Example 1 for the preparation ofCBZ-L-Leu-L-Phe-L-methioninol, this derivative was synthesized fromFmoc-L-4-Me-Phe (purchased from Neosystems Laboratories, Strasbourg,France) and L-(Tr-glutaminol) in 85% yield and isolated as a whitesolid. IR (KBr) 3316, 3283, 3024, 2946, 1694, 1667, 1448, 1256, 1041,760, 700 cm⁻¹; ¹H NMR (DMSO-d₆) δ 1.56 (m, 1H), 177 (m, 1H), 222 (s,3H), 2.26 (m, 2H), 2.74 (m, 1H), 2.90 (m, 1H), 3.17 (m, 1H), 3.69 (m,1H), 4.03-4.23 (m, 4H), 7.03-7.54 (m, 21H), 7.39 (t, 2H, J=7.4 Hz), 7.50(d, 1H, J=8.5 Hz), 7.59 (d, 1H, J=7.4 Hz), 7.60 (d, 1H, J=7.7 Hz), 7.70(d, 1H, J=8.8 Hz), 7.87 (d, 2H, J=7.4 Hz), 8.45 (s, 1H); MS calcd forC₄₉H₄₇N₃O₅+Cs 890, found 890.

Prepartion of Intermediate L-(4-Me-(Phe)-L-(Tr-Glutaminol)

To a solution of Fmoc-L-(4-Me-Phe)-L-(Tr-glutaminol) (3.25 g, 4.29 mmol)in anh DMF (10 mL) was added piperidine (0.51 mL, 5.15 mmol). Thesolution was stirred and monitored by TLC. Upon consumption of thestarting material, the reaction mixture was concentrated to a residueand then subjected to column chromatography on silica gel (5%MeOH/CH₂Cl₂) to afford the product as white solid in 87% yield. IR (KBr)3326, 3054, 3030, 2953, 2872, 1651, 1516, 1491, 1447, 1036, 700 cm⁻¹; ¹HNMR (DMSO-d₆) δ 1.47 (m, 1H), 1.75 (m, 3H), 2.13 (m, 1H), 2.23 (s, 3H)2.57 (dd, 1H, J=13.2, 8.1 Hz), 2.88 (dd, 1H, J=13.6, 4.8 Hz), 3.20 (m,1H), 3.30 (m, 1H), 3.66 (m, 1H), 4.64 (t, 1H, J=5.5 Hz), 7.07 (m, 4H),7.10-7.28 (m, 15H), 7.62 (d, 1H, J=8.8 Hz), 8.54 (s, 1H); MS calcd forC₃₄N₃₇N₃O₃+Na 558, found 558.

Preparation of Intermediate Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)

A stirred suspension of L-α-(t-butyl-Gly) (0.656 g, 5.0 mol) in 18 mLCH₂Cl₂, and diisopropylethylamine (3.5 mL, 20 mmol) was cooled to 0° C.To this mixture chlorotrimethylsilane (0.83 mL, 6.5 mmol) was addeddropwise. The slurry was allowed to warm to rt, and the mixture wasstirred for about 2 h. At this time the mixture was recooled to 0° C.,and cyclopentyl chlorothiolformate (0.823 g, 5.0 mmol) was addeddropwise. The slurry became a pale yellow solution after stirring at rtfor approximately 5 h. The solution was concentrated, redissolved in anexcess of EtOAc and washed with H₂O, 10% aq KHSO₄, H₂O and brine. Theorganic phase was dried over MgSO₄, filtered and concentrated to givecyclopentylthiocarbonyl-L-α-(t-butyl-Gly) as a yellow oil in nearlyquantitative yield which was azetroped with benzene to remove anyresidual water before being used in the next step. IR (film) 3324, 2965,2920, 2872, 1726, 1642, 1518, 1202 cm⁻¹; ¹H NMR (CDCl₃) δ 1.03 (s, 9H),1.48-1.73 (m, 6H), 2.10 (m, 2H), 3.72 (m, 1H), 4.46 (m, 1H), 5.79(m,1H); MS calcd for C₁₂H₂₁NO₃S+Na 282, found 282.

Preparation of IntermediateCyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Glutaminol)

This preparation was carried out following the procedure of L. A.Carpino, J. Am. Chem. Soc. 1993, 115, 4397.Cyclopentylthiocarbonyl-L-α-(t-butyl-Gly) (0.325 g, 1.25 mmol) wasdissolved in 8.0 mL of DMF. Diisopropylethylamine (0.45 mL, 2.5 mmol)was added, followed by 0.67 g (1.25 mmol) ofN-Me-L-(4-Me-Phe)-L-(Tr-glutaminol). The reaction was cooled to 0° C.and O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexaflurophosphate (HATU) (0.476 g, 1.25 mmol) was added. The reactionmixture was allowed to warm to rt whereupon the DMF was removed invacuo. The residue was dissolved with EtOAc, and the organic phasewashed consecutively with 1N HCl, a sat NaHCO₃ solution, H₂O, and brine.The solvent was dried over MgSO₄, filtered, and concentrated to give aresidue which was subjected to column chromatography on silica gel(gradient; 2-5% MeOH/CHCl₃) to give 0.95 g (98%) of a white amorphoussolid: IR(KBr) 3302, 2957, 2876, 1669, 1645, 1537, 1447, 1196, 700 cm⁻¹;¹H NMR (CDCl₃) δ 0.88 (s, 9H), 1.48-170 (m, 9H), 1.85 (m, 1H), 2.04 (m,2H), 2.28 (s, 3H), 2.32 (m, 2H), 2.92 (m, 2H), 3.25 (dd, 1H, J=8.1, 3.5Hz), 3.30 (dd, 1H, J=10.9, 3.7 Hz), 3.66 (m, 1H), 3.72 (m, 1H, 4.14 (m,1H), 4.47 (m, 1H), 6.04 (d, 1H, J=7.7 Hz), 6.52 (d, 1H, J=7.7 Hz), 6.60(d, 1H, J=7.0 Hz), 7.05 (m, 5H), 7.24 (m, 15H), MS calcd forC₄₆H₅₆N₄O₅S+Na 799, found 799.

Preparation of IntermediateCyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Glutaminal)

Using the general procedure described in Example 1 for the preparationof CBZ-L-Leu-L-Phe-L-methioninal (sulfoxide),cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminal)was synthesized fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminol)in quantitative yield and isolated as a white amorphous solid and usedwithout further purification: IR(film) 3302, 3061, 3030, 2961, 2870,1730, 1644, 1514, 1493, 1196, 911, 733, 700 cm⁻¹; ¹H NMR (CDCl₃) δ 0.90(s, 9H), 1.46-1.68 (m, 8H), 1.86 (m, 1H), 2.00-2.24 (m, 2H), 2.28 (s,3H), 2.31 (m, 1H), 2.96 (m, 2H), 3.58 (m, 1H), 4.05 (m, 1H), 4.14 (m,1H), 4.52 (m, 1H), 5.88 (m, 1H), 6.28 (m, 1H), 6.90 (m, 1H), 7.07 (m,5H), 7.25 (m, 15H), 9.30 (s, 1H); MS calcd for C₄₆H₅₄N₄O₅S•CH₃OH(methyl-hemiacetal)+Na 829, found 829.

Preparation of IntermediateEthyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide)]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-propenoatewas synthesized fromcyclopentlythiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminal)(0.468 g, 0.627 mmol) to give 0.52 g of material contaminated withtriphenylphosphine oxide after column chromatography on silica gel(gradient: 1-2.5% MeOH/CH₂Cl₂), which was used without furtherpurificaation: white amorphous solid: IR(film) 3302, 3061, 2967, 2868,1721, 1642, 1514, 1491, 1370, 1192, 1036, 911, 731, 700 cm⁻¹; ¹H NMR(CDCl₃) δ 0.72 (s, 9H), 1.29 (t, 3H, J=7.0 Hz), 1.46-1.68 (m, 6H),1.86-2.05 (m, 4H), 2.29 (s, 3H), 2.32 (m, 2H), 2.91 (m, 2H), 3.00 (m,1H), 3.62 (m, 1H), 4.07 (m, 1H), 4.17 (q, 2H, J=7.2 Hz), 4.43 (m, 2H),5.61 (dd, 1H, J=15.8, 1.5 Hz), 5.95 (m, 1H), 6.34 (m, 1H), 6.57 (m, 1H),6.64 (dd, 1H, J=15.8, 5.5 Hz), 7.03 (m, 5H), 7.24 (m, 15H), MS calcd forC₅₀H₆₀N₄O₆S+Na 867, found 867.

Preparation ofProduct-Ethyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the preparation ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-propenoatewas synthesized fromethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-propenoateand isolated as a white solid after purification by columnchromatography on silicia gel using a gradient solvent system (1-5%MeOH/CH₂Cl₂) (57% yield; two steps fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminal):IR (KBr) 3318, 2973, 2951, 2868, 1715, 1651, 1539, 1371, 1192 cm ⁻¹; ¹HNMR (DMSO-d₆) δ 0.83 (s, 9H), 1.21 (t, 3H, J=7.2 Hz), 1.41-1.72 (m, 8H),2.02 (m, 4H), 2.22 (s, 3H), 2.81 (m, 2H), 3.54 (m, 1H), 4.10 (q, 2H,J=7.0 Hz), 4.24 (d, 1H, J=9.3 Hz), 4.36 (m, 1H), 4.43 (m, 1H), 5.56 (dd,1H, J=15.7, 1.4 Hz), 6.65 (dd, 1H, J=15.7, 5.5 Hz), 6.73 (s, 1H), 7.03(m, 4H), 7.13 (s, 1H), 7.86 (d, 1H, J=9.3 Hz), 8.04 (d, 1H, J=8.4 Hz),8.12 (d, 1H, J=7.8 Hz); HRMS calcd for C₃₁H₄₆N₄O₆S+Cs 735.2192, found735.2180. Anal. (C₃₁H₄₆N₄O₆S) C, H, N, S.

Example 39 Preparation of Compound 175:Ethyl-2-Methyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-PropenoatePreparation of IntermediateEthyl-2-Methyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide)]-E-propenoate,ethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-propenoatewas synthesized fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminal)(0.466 g, 0.60 mmol) and (carbethoxyethylidene)tripentylphosphorane(0.24 g, 0.66 mmol) to give 0.487 g of material contaminated withtriphenylphosphine oxide after column chromatography on silica gel(gradient: 1-2.5% MeOH/CH₂Cl₂) which was used without furtherpurification. white amorphous solid: IR(film) 3302, 3063, 2967, 2870,1711, 1642, 1516, 1491, 1250, 1194, 911, 731, 698 cm ⁻¹; ¹H NMR (CDCl₃)δ 0.88 (s, 9H), 1.31 (t, 3H, J=7.2 Hz), 1.50-1.77 (m, 6H), 1.81 (m, 2H),1.82 (s, 3H), 2.06 (m, 2H), 2.28 (s, 3H), 2.31 (m, 2H), 2.93 (m, 2H),3.64 (m, 1H), 4.04 (m, 1h), 4.20 (q, 2H, J=7.0 Hz), 4.40 (m, 1H), 4.58(m, 1H), 5.90 (m, 1H), 6.30 (m, 3H), 7.01 (m, 5H), 7.24 (m, 15H). MScalcd for C₅₁H₆₂N₄O₆S+Na 881, found 881.

Preparation ofProduct-Ethyl-2-Methyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the preparation ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-Gln]-E-propenoatewas synthesized fromethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-Gln)]-E-propenoateand isolated as a white solid after purification by columnchromatography on silica gel using a gradient solvent system (1-5%MeOH/CH₂Cl₂) (55% yield; two steps fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminal):IR (KBr) 3324, 2963, 2870, 1707, 1647, 1550, 1516, 1257, 1196 cm⁻¹; ¹HNMR (DMSO-d₆) δ 0.83 (s, 9H), 1.22 (t, 3H, J=7.2 Hz), 1.41-1.73 (m, 8H),1.77 (m, 3H), 2.00 (m, 4H), 2.20 (s, 3H), 2.78 (m, 2H), 3.55 (m, 1H),4.12 (q, 2H, J=7.0 Hz), 4.23 (d, 1H, J=9.0 Hz), 4.35 (m, 1H), 4.48 (m,1H, ), 6.29 (dd, 1H, J=9.3, 1.2 Hz), 6.72 (s, 1H), 6.99 (m, 4H), 7.13(s, 1H), 7.86 (d, 1H, J=9.0 Hz), 8.03 (m, 2H); HRMS calcd forC₃₂H₄₈N₄O₆S+Cs 749.2349, found 749.2336. Anal. (C₃₂H₄₈N₄O₆S) C, H, N, S.

Example 40 Preparation of Compuond 176:Ethyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-PropenoatePreparation of IntermediateCyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-(Tr-Glutaminol)

This intermediate was prepared as a white solid in 75% yield fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly) and the free base ofL-(4-F-Phe)-L-(Tr-glutaminol) HCl using the procedure described topreparecyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-Me-Phe)-L-(Tr-glutaminol).IR (KBr) 3299, 3063, 2969, 2870, 1651, 1510, 1447, 1225, 1192, 766, 700cm⁻¹; ¹H NMR (CDCl₃) δ 0.88 (s, 9H), 1.50-1.76 (m, 9H), 1.85 (m, 1H),2.05 (m, 2H), 2.36 (m, 2H), 2.50 (m, 1H), 2.92 (m, 2H), 3.32 (m, 2H),3.66 (m, 1H), 3.73 (m, 1H), 4.17 (m, 1H), 4.69 (m, 1H), 6.09 (d, 1H,J=7.0 Hz), 6.74 (m, 1H), 6.91 (m, 2H), 7.05 (m, 2H), 7.24 (m, 15H). MScalcd for C₄₆H₅₃N₄O₅SF+Na 803, found 803.

Preparation of IntermediateCyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-(Tr-Glutaminal)

Using the general procedure described in Example 1 for the preparationCBZ-L-Leu-L-Phe-L-methioninal (sulfoxide),cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminal)was synthesized fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminol)in quantitative yield and isolated as a white amorphous solid and usedwithout further purification: IR (film) 3302, 3061, 3030, 2961, 2866,1732, 1644, 1510, 1447, 1225, 1196, 911, 733, 700 cm⁻¹; ¹H NMR (CDCl₃) δ0.90 (s, 9H), 1.48-1.67 (m, 8H), 1.85 (m, 1H), 2.00-2.28 (m, 2H), 2.63(m, 2H), 2.90 (dd, 1H, J=14.9, 6.1 Hz), 3.03 (dd, 1H, J=14.5, 6.8 Hz),3.64 (m, 1H), 4.07 (m, 1H), 4.18 (m, 1H), 4.53 (m, 1H), 5.92 (m, 1H),6.31 (m, 1H), 6.92 (m, 2H), 7.10 (m, 3H), 7.23 (m, 15H), 9.31 (s, 1H);MS calcd for C₄₅H₅₃N₄O₅SF•CH₃OH (methyl-hemiacetal)+Na 833, found 833.

Preparation of IntermediateEthyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-(Tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide)]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-Gln)]-E-propenoatewas synthesized fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminal)(0.343 g, 0.44 mmol) to give 0.377 g of material contaminated withtriphenylphosphine oxide after column chromatography on silica gel(gradient: 1-2.5% MeOH/CH₂Cl₂) which was used without furtherpurification: white amorphous solid: IR (KBr) 3314, 3285, 2969, 2936,1723, 1651, 1510, 1447, 1370, 1190, 1038, 700 cm⁻¹; ¹H NMR (CDCl₃) δ0.88 (s, 9H), 1.28 (t, 3H, J=7.0 Hz), 1.48-1.78 (m, 8H), 1.83-2.15 (m,4H), 2.32 (m, 2H), 2.85 (m, 1H), 3.00 (m, 1H), 3.61 (m, 1H), 4.16 (q,2H, J=7.0 Hz), 4.39 (m, 2H), 5.54 (d, 1H, J=15.4 Hz), 6.17 (m, 1H), 6.63(dd, 1H, J=15.4, 4.0 Hz), 6.91 (m, 2H), 7.01 (m, 2H), 7.28 (m, 15H),7.45 (m, 1H), 7.54 (m, 1H), 7.63 (m, 1H). MS calcd for C₄₉H₅₇N₄O₆SF+Na871, found 871.

Preparation ofProduct-Ethyl-3-[cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the preparation ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-propenoatewas synthesized fromethyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-Gln)]-E-propenoateand isolated as a white solid after purification by columnchromatography on silica gel using a gradient solvent system (1-5%MeOH/CH₂Cl₂) (56% yield; two steps fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminal):IR (KBr) 3310, 2961, 2868, 1713, 1649, 1512, 1192 cm⁻¹; ¹H NMR (DMSO-d₆)δ 0.83 (d, 9H), 1.21 (t, 3H, J=7.2 Hz), 1.40-1.69 (m, 8H), 2.01 (m, 4H),2.80 (dd, 1H, J=14.0, 8.1 Hz), 2.90 (dd, 1H, J=13.2, 7.0 Hz), 3.54(quin, 1H, J=7.2 Hz), 4.09 (q, 2H, J=6.9 Hz), 4.28 (d, 1H, J=9.6 Hz),4.38 (m, 1H), 4.47 (m, 1H), 5.48 (dd, 1H, J=15.6, 1.3 Hz), 6.64 (dd, 1H,J=15.6, 5.3 Hz), 6.74 (bs, 1H), 7.00 (t, 2H, J=8.8 Hz), 7.13 (bs, 1H),7.20 (d, 1H, J=8.5 Hz), 7.22 (d, 1H, J=8.5 Hz), 7.88 (d, 1H, J=9.2 Hz),8.08 (d, 1H, J=8.1 Hz), 8.18 (d, 1H, J=7.7 Hz); HRMS calcd forC₃₀H₄₃N₄O₆SF+Cs 739.1942, found 739.1954. Anal. (C₃₀H₄₃N₄O₆SF) C, H, N,S.

Example 41 Preparation of Compound 177:Ethyl-2-Methyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-PropenoatePreparation of IntermediateEthyl-2-Methyl-3-[Cyclopentylthicarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-(tr-Gln)]-E-Propenoate

Using the procedure described in Example 1 for the preparation ofethyl-3-[CBZ-L-Leu-L-Phe-L-Met (sulfoxide)]-E-propenoate,ethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-Gln)]-E-propenoatewas synthesized fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminal)(0.297 g, 0.38 mmol) and (carbethoxyethylidene)triphenylphosphorane(0.152 g, 0.42 mmol) to give 0.377 g of material contaminated withtriphenylphosphine oxide after column chromatography on silica gel(gradient: 1-2.5% MeOH/CH₂Cl₂) which was used without furtherpurification. white amorphous solid: IR(film) 3356, 3291, 3063, 2973,2951, 1711, 1651, 1510, 1447, 1256, 1190, 752, 700 cm⁻¹; ¹H NMR (CDCl₃)δ 0.90 (s, 9H), 1.31 (t, 3H, J=7.0 Hz), 1.51-1.83 (m, 11H), 2.17 (m,2H), 2.28 (m, 2H) 2.75-3.02 (m, 2H), 3.66 (m, 1H), 4.16 (m, 3H), 4.45(m, 1H), 4.60 (m, 1H), 6.30 (m, 2H) 6.58 (m, 1H), 6.78 (m, 1H), 6.88 (m,2H), 6.98 (m, 3H), 7.20 (m, 15H). MS calcd for C₅₀H₅₉N₄O₆SF+Na 885,found 885.

Preparation ofProduct-Ethyl-2-Methyl-3-[Cyclopentylthiocarbonyl-L-α-(t-Butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-Propenoate

Using the procedure described in Example 34 for the preparation ofethyl-3-[ethylthiocarbonyl-L-α-(t-butyl-Gly)-L-Phe-L-Gln]-E-propenoate,ethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-Gln]-E-propenoatewas synthesized fromethyl-2-methyl-3-[cyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-Gln)]-E-propenoateand isolated as a white solid after purification by columnchromatography on silica gel using a gradient solvent system (1-5%MeOH/CH₂Cl₂) (55% yield; two steps fromcyclopentylthiocarbonyl-L-α-(t-butyl-Gly)-L-(4-F-Phe)-L-(Tr-glutaminal):IR (KBr) 3326, 2951, 2868, 1713, 1645, 1533, 1510, 1260, 1194 cm⁻¹; ¹HNMR (DMSO-d₆) δ 0.83 (s, 9H), 1.22 (t, 3H, J=7.0 Hz), 1.41-1.75 (m, 8H),1.77 (m, 3H), 1.92 (m, 4H), 2.77 (dd, 1H, J=13.8, 8.3 Hz), 2.85 ((dd,1H, J=13.6, 7.0 Hz), 3.55 (quin, 1H, J=7.0 Hz), 4.12 (q, 2H, J=7.1 Hz),4.22 (d, 1H, J=9.2 Hz), 4.38 (m, 1H), 4.45 (m, 1H), 6.24 (dd, 1H, J=9.2,1.5 Hz), 6.72 (bs, 1H), 6.96 (t, 2H, J=8.8 Hz), 7.87 (d, 1H, J=8.8 Hz),8.03 (d, 1H, J=8.1 Hz), 8.11 (d, 1H, J=7.7 Hz); HRMS calcd forC₃₁H₄₅N₄O₆SF+Cs 753.2098, found 753.2084. Anal. (C₃₁H₄₅N₄O₆SF) C, H, N,S.

Example 42 Preparation of Compound 179:Ethyl-3-(Cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-Gln)-E-PropenoatePreparaion of ProductEthyl-3-(Cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-Gln)-E-Propenoate

Using the procedure described in Example 36 for the preparation ofethyl-3-[cyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-Gln]-E-propenote,ethyl-3-(cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-Gln)-E-propenoatewas synthesized fromethyl-3-(cyclopentylthiocarbonyl-L-[S(O)-Me-Pen]-L-Phe-L-[Tr-Gln])-E-propenoatein 40% yield (3 steps fromcyclopentylthiocarbonyl-L-(S-Me-Pen)-L-Phe-L-(Tr-glutaminol)): whiteamorphous solid: IR (KBr) 3302, 1662, 1541, 1458, 1205, 1138, 1028 cm⁻¹;¹H NMR (DMSO-d₆) (mixture of diastereomers) δ 1.03 (s), 1.12 (s), 1.21(t, 3H, J=6.0 Hz), 1.42-1.76 (m), 2.0-2.21 (m), 2.34 (s), 2.42 (s),2.80-2.87 (m), 2.93-3.11 (m), 3.47-3.60 (m), 4.10 (q, J=6.0 Hz),4.35-4.40 (m), 4.44-4.52 (m), 4.64 (d, J=6.0 Hz), 5.58-5.62 (m),6.60-6.70 (m), 6.75 (bs), 7.14-7.21 (m), 8.16-8.22 (m), 8.41 (d, J=9.0Hz), 8.54 (d, J=9.0 Hz). HRMS calcd for C₃₀H₄₄N₄O₇S₂+Cs 769.1706, found769.1727.

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modification can be made therein withoutdeparting from the spirit and scope thereof. Thus, it is intended thatthe present invention cover the modifications and variations, providedthey come within the scope of the appended claims and their equivalents.

BIOCHEMICAL AND BIOLOGICAL EVALUATION Inhibition of Rhinovirus Protease

Stock solution (50 mM, in DMSO) of various compounds were prepared;dilutions were in the same solvent. Recombinant Rhinovirus 3C proteasesfrom serotypes 14, 16, 2 or 89 were prepared by the following standardchromatographic procedures: (1) ion exchange using Q Sepharose Fast Flowfrom Pharmacia; (2) affinity chromatography using Affi-Gel Blue fromBiorad; and (3) sizing using Sephadex G-100 from Pharmacia. Assayscontained 2% DMSO, 50 mM tris pH 7.6, 1 mM EDTA, a compound at theindicated concentrations, approximately 1 μM substrate, and 50-100 nMprotease. For K_(i) determinations, the compound and the enzyme werepreincubated for 10 minutes at 30° C. prior to addition of the substrate(substrate start). The k_(obs/I) values were obtained from reactionsinitiated by addition of enzyme rather than substrate. RVP activity ismeasured in the fluorescence resonance energy transfer assay. Thesubstrate was (N-terminal)DABCYL-(Gly-Arg-Ala-Val-Phe-Gln-Gly-Pro-Val-Gyl)-EDANS. In the uncleavedpeptide, the EDANS fluorescence was quenched by the proximal DABCYLmoeity. When the peptide was cleaved, the quenching was relieved, andactivity was measured as an increase in fluorescence signal. Data wasanalyzed using standard non linear fitting programs (Enzfit), and areshown Table 1.

TABLE 1 COMPOUND # RVP INHIB k_(obs/I) (M-1 sec-1) 1 77 (50) ND 2 6.6μM(K_(i)) ND 3 81(0.1) 37,000 (16) 6,500 (89) 3,400 (2) 1,900 4 49(0.5)790 5 7.1 μM(K_(i)) 221 6 32 μM(K_(i)) 350 7 9.5 μM(K_(i)) 2,400 (16)42(1) ND 8 36 μM(K_(i)) 61 9 20(1) 160 10 55(5) 270 11 28 μM(K_(i))20,000 12 4.3 μM(K_(i)) 2,200 13 6.5 μM(K_(i)) 54,000 (16) 9,000 (2)2,400 (89) 5,500 14 NI ND 15 55(50) 27 16 40(0.25) 3,500 17 1.25μM(K_(i)) 6,100 18 15.3 μM(K_(i)) 7,700 19 35 μM(K_(i)) 7,900 20 NI ND21 9.9 μM(K_(i)) 2,100 22 4.3 μM(K_(i)) 1,300 23 177 μM(K_(i)) 120 24 ND500,000 25 5.5 μM(K_(i)) 3,700 26 52(0.1) 5,400 27 20 μM(K_(i)) 3,000 2857 μM(K_(i)) 4,000 29 ND ND 30 373 μM(K_(i)) 430 31 25(10) 21 32 ND 28033 24(10) 33 34 10(10) 34 35 16.5 μM(K_(i)) 46,388 (2) ND 2,357 (16) ND9,177 36 15 μM(K_(i)) 12,000 37 18.8 μM(K_(i)) 5,900 38 >50 μM(K_(i))400 39 ND 1,200 40 ND 250 41 ND 8,464 42 ND 150,000 43 ND 4,500 44 12.6μM(K_(i)) 21,000 45 NI ND 46 ND 120,000 49 ND 460,000 51 ND 310,000 52ND 15,000 53 ND 11,320 56 15 μM(K_(i)) 5,624 59 2.0 μM(K_(i)) 200 60 5.0μM(K_(i)) 575 61 ND 125,940 (2) ND 14,000 (16) ND 25,000 62 ND 600,000(2) ND 600,000 (16) ND 300,000 65 2.9 μM(K_(i)) ND 66 ND 400,000 67 ND9,600 68 15 μM(K_(i)) 750 70 ND 39,000 71 ND 20,650 73 ND 20,000 (2) ND1,750 (16) ND 4,500 74 2.4 μM(K_(i)) 75 ND 76 30 μM(K_(i)) ND 77 4.8μM(K_(i)) ND 78 7.0 μM(K_(i)) 79 ND 13,900 80 ND 200,000 81 ND 124,00082 26 μM(K_(i)) 7,300 83 8.0 μM(K_(i)) ND 84 ND 18,650 85 3.0 μM(K_(i))6,500 86 4.0 μM(K_(i)) 12,000 87 6.0 μM(K_(i)) 5,430 88 >30 μM(K_(i))8,960 89 5 μM(K_(i)) 53,360 (16) ND 2,800 90 ND 10,918 (16) ND 3,600 9110 μM(K_(i)) 5,427 92 ND 445 93 30 μM(K_(i)) 3,444 94 1.5 μM(K_(i))5,800 95 ND <1000 96 ND 300 97 ND 12,900 98 ND 91 99 10 (50) ND 100 ND1,200 101 ND 11,288 102 12 μM(K_(i)) 3,845 103 ND 29,200 (2) ND 1,106(16) ND 3,354 104 2.5 μM(K_(i)) 8,000 (16) 1.5 (μM(K_(i)) ND 105 ND1,200 106 2.0 μM(K_(i)) 280,000 (2) ND 28,400 (16) ND 75,000 107 13.5μM(K_(i)) 3,655 108 ND 4,694 109 ND 1,348 110 ND 9,072 111 5.0 μM(K_(i))2,065 112 13 μM(K_(i)) 6,800 113 ND 8,877 114 ≧1.0 μM(K_(i)) 82,320 (2)ND 1,971 115 11 μM(K_(i)) 4,485 116 ND 23,670 117 ND 18,760 118 39μM(K_(i)) 1,448 119 5.0 μM(K_(i)) 69,800 120 6.0 μM(K_(i)) 91,300 (2) ND8,900 (16) ND 20,034 121 12 μM(K_(i)) 238 122 ND 1,252 123 ND 890 124 ND1,000 125 ND >500,000 126 ND 29,000 127 ND 28,347 128 ND 22,691 129 ND230,000 130 30-40 nM(K_(i)) ND 131 NI NI 132 10 μM(K_(i)) 10,800 133 ND9,600 134 ND 1,769 135 ND 16,270 (2) ND 671 (16) ND 3,465 136 ND 4,210137 ND 2,344 (2) ND 643 (16) ND 1,157 138 20 μM(K_(i)) 1,769 139 ND43,140 (2) ND 691 (16) ND 1,259 140 ND 7,122 141 ND 2,309 142 ND 2,929143 ND 2,963 144 ND ND 145 10-20 μM(K_(i)) ND 146 ND 62,500 (2) ND 7,790(16) ND 16,900 147 ND 18,600 (2) ND 1,000 (16) ND 4,290 148 1.0μM(K_(i)) 57,000 (2) ND 8,300 (16) ND 14,800 149 ND 39,940 (2) ND 2,840(16) ND 7,700 150 ND 573 151 >4.8 μM(K_(i)) 39,750 152 3.2 μM(K_(i))38,900 153 1.4 μM(K_(i)) 141,200 (2) ND 13,350 (16) ND 30,650 154 1.1μM(K_(i)) 78,900 (2) ND 5,400 (16) ND 13,900 155 4.2 mM(K_(i)) 59,425(2) ND 1,390 (16) ND 5,250 156 ND NI 157 6.0 μM(K_(i)) 161,500 (2) ND9,700 (16) ND 30,800 158 17 μM(K_(i)) 22,600 (2) ND 2,200 (16) ND 6,400159 0.5 μM(K_(i)) 35,000 (2) ND 2,500 (16) ND 6,500 160 ND 312,000 (2)ND 26,710 (16) ND 50,000 161 ND 1,086,000 (2) ND 200,000 (16) ND 126,000162 ND 800,000 (2) ND 150,000 (16) ND 80,000 163 3.6 μM(K_(i)) 9,800 164ND 155,500 165 ND 97,000 (2) ND 5,600 (16) ND 20,200 166 ND 40,900 (2)ND 3,500 (16) ND 7,700 167 ND 165,400 (2) ND 10,700 (16) ND 42,100 168ND 37,800 169 ND 800 170 ND 85,300 (2) ND 8,400 (16) ND 30,000 171 ND21,200 (2) ND 830 (16) ND 3,250 172 ND 31,700 (2) ND 2,000 (16) ND 6,000173 ND 1,000,000 (2) ND 113,000 (16) ND 185,000 174 ND 800,000 175 ND124,000 176 0.48 μM(K_(i)) 240,000 177 ND 80,300 178 ND 286,300 179 0.36μM(K_(i)) 300,000 180 0.42 μM(K_(i)) 300,000 181 ND 1,000,000 182 ND114,360 183 0.55 μM(K_(i)) 500,000 (16) ND 60,000 184 ND 59,900 185 ND600,000 186 ND 950,000 187 NI ND 188 0.16 μM(K_(i)) 580,000 189 ND386,000 190 ND 29,230

In the above table, all data is RVP serotype-14 unless otherwise notedin parentheses. All strains of human rhinovirus (HRV) were purchasedfrom American Type Culture Collection (ATCC), except for serotype 14,which was produced from the infectious cDNA clone constructed andsupplied to us by Dr. Roland Rueckert at the Institute for MolecularVirology, University of Wisconsin, Madison, Wisc. The column designatedINHIB represents the percent inhibition, with the concentration of thecompound in μM indicated in parentheses, unless K_(i) was assigned asdesignated by (K_(i)), at 10 minute preincubation with 50 nM RVP priorto addition of substrate was used. The data in the column designatedk_(obs/I) was measured from progress curves in enzyme start experiments.The designation NI indicates that no inhibition was obtained when 10 μMof a compound was used. The designation ND indicates that a value wasnot determined for that compound.

Antirhinoviral HI-HeLa Cell Culture Assay

In the Cell Protection Assay, the ability of compounds to protect cellsagainst HRV infection was measured by the XTT dye reduction method. Thismethod is described in Weislow, O. S., R. Kiser, D. L. Fine, J. Bader,R. H. Shoemaker, and M. R. Boyd, J. Natl. Cancer Inst. 1989, 81,577-586, which is incorporated herein by reference.

HI-HeLa cells were infected with HRV-14 at a multiplicity of infection(m.o.i.) of 0.13 (virus particles/cell) or mock-infected with mediumonly. Infected or mock-infected cells were resuspended at 8×10⁵ cellsper mL and incubated with appropriate concentration of compounds offormulas I and II. Two days later, XTT/PMS was added to test plates andthe amount of formazan produced was quantified spectrophotometrically at450/650 nm. The EC₅₀ was calculated as the concentration of compoundthat increased the percentage of formazan production incompound-treated, virus-infected cells to 50% of that produced bycompound-free mock-infected cells. The 50% cytotoxic dose (CC₅₀) wascalculated as the concentraion of compound that described the percentageof formazan produced in compound-treated, mock-infected cells to 50% ofthat produced in compound-free, mock-infected cells. The therapeuticindex (TI) was calculated by dividing the CC₅₀ by the EC₅₀.

All strains of human rhinovirus (HRV) for use in this assay werepurchased from American Type Culture Collection (ATCC), except for HRVserotype-14, which was produced from the infectious cDNA clone,constructed and supplied to us by Dr. Roland Rueckert at the Instituefor Molecular Virology, University of Wisconsin, Madison, Wisc. HRVstocks were propagated, and antiviral assays were perforemd in HI-HeLacells (ATCC). Cells were grwon in Minimal Essential Medium, availablefrom Life Technologies, with 10% fetal bovine serum.

The compounds were tested against control compounds WIN 51711, WIN52084, and WIN 54954, all obtained from Sterling-WinthropPharmaceuticals, and control compound Pirodavir, obtained from JanssenPharmaceuticals.

TABLE 2 Compound # EC₅₀ (μM) CC₅₀ (μM) TI 1 ND ND 2 100 >320 >3.2 30.61 >320 >525 4 2.2 >320 >146 5 1.6 251 157 6 >320 >320 7 3.2 >320 >1008 >320 >320 >5 9 >320 >320 10 200 >320 >2 11 1.3 >320 >246 121.6 >100 >63 13 2.0 58.9 29 14 17.8 500 28 15 >100 >100 16 32 >100 >3 171.8 >100 >56 18 0.64 >100 >156 19 1.35 >100 >74 20 >320 >320 2122.4 >100 >5 22 56.2 251 >5 23 >100 >100 24 4.0 16 4 25 3.1 >100 >33 262.0 44.7 22 27 3.5 160 46 28 4.5 63.1 14 29 27 500 19 30 5.6 100 18 3150.1 >100 >2 32 10 >100 >10 33 794 >100 >1 34 100 >100 >1 351.8 >320 >178 36 5.6 >320 >57 37 4.0 >100 >25 38 >320 >320 39 >320 >32040 >100 >100 41 56 56 1 42 22.4 100 >4 43 10 18 44 1.0 >320 >32045 >100 >100 46 3.2 45 14 49 2.4 19.1 8 51 32 32 52 1.7 5.6 3 535.3 >320 >60 56 1.6 >320 >203 59 >320 >320 60 158 >320 >2 61 0.89 56 6362 1.6 >100 >63 65 158 >320 >2 66 1.4 6.3 5 67 5.2 >320 >62 6816 >320 >20 70 1.2 >320 >267 71 14.1 >320 23 73 ND 74 10 250 25 755.0 >100 >20 76 >320 >320 77 >320 >320 78 10 79.4 8 79 45 >320 >7 8050 >320 >6 81 8.0 112 14 82 3.0 >320 >107 83 100 >320 >3 84 16 >320 >2085 16 >320 >20 86 17 >320 >19 87 10.6 >320 >30 88 8.8 >160 >18 89 1.8 2916 90 5.2 >320 >61 91 56 >320 >6 92 5.6 56 10 93 >320 >320 9446.8 >320 >7 95 >320 >320 96 19.1 100 5 97 >320 >320 98 100 >320 >3.2 99141 >320 >2 100 11.1 >320 >29 101 2.0 >320 >160 102 5.6 >320 >57 1031.7 >320 >188 104 5.2 >320 >61 105 14 >320 >23 106 0.27 >320 >1185 10713.5 >320 >23 108 6.0 >320 >53 109 20 >320 >16 110 1.3 >320 >246 11129.5 >320 >11 112 27 >320 >12 113 10 >320 >32 114 0.55 >320 >582 11519 >320 >17 116 0.6 >320 >533 117 1.0 >320 >320 118 17.8 >320 >18 1191.1 >320 >291 120 0.46 >320 >695 121 >320 >320 122 1.78 10 5123 >320 >320 124 126 >320 >2 125 >100 100 126 >320 >320 127 >100 ND128 >320 >320 129 >320 >320 130 15.8 >100 >6 131 >100 >100 1325.6 >320 >57 133 >177 177 134 56.2 >320 >5 135 1.9 >320 >168136 >320 >320 137 223.9 >320 >1 138 >41.7 41.7 139 3.5 >320 >91 14039 >320 >8 141 5.4 >320 >59 142 8.9 >320 >36 143 10 >320 >32 144103.5 >320 >3 145 >320 >320 146 0.38 >320 >842 147 205 >320 1480.25 >320 >1280 149 1.78 >320 >180 150 >320 >320 151 0.32 177.8 555 1521.78 >320 >180 153 0.12 >320 >2667 154 5.5 >320 >58 155 0.18 >320 >1778156 35.5 >320 >9 157 0.56 >320 >571 158 5.9 >320 >54 159 2.4 >320 >133160 5.0 >320 >64 161 0.17 >100 >588 162 0.32 >100 >312 163 0.5 >100 >200164 0.71 >100 >141 165 0.20 >100 >500 166 5.6 >100 >18 1670.083 >100 >1204 168 0.32 >100 >312 169 18 >100 >5 170 0.20 >100 >500171 0.71 >100 >140 172 0.79 >100 >126 173 0.08 >100 >1250 1740.056 >100 >1786 175 0.18 >100 >555 176 0.14 >100 >714 177 0.5 >100 >200178 0.10 >100 >1000 179 1.78 >100 >56 180 0.056 >100 >1785 1810.1 >100 >1000 182 0.18 >100 >556 183 0.03 >100 >3333 184 0.19 >100 >526185 0.50 >100 >200 186 ND ND 187 ND ND 188 ND ND WIN 51711 0.78 >60 >77WIN 52084 0.07 >10 >143 WIN 54954 2.13 >63 >30 Pirodavir 0.03 >10 >300

Normal Human Bronchial Epithelial Cell Assay

Normal human bronchial cells were obtained from cadavers and cultured.The cells were plated at 2×10⁴ per well in a 86 well plate. They wereallowed to adhere and grow for 24 hours in 200 μL of serum-freebronchial/tracheal epithelial cell growth medium at 37° C. with 5% CO₂.Human Rhinovirus-serotype 10 (HRV-10) was purchased from American TypeCulture Collection (ATCC). To start the assay, the supernatant wasremoved, and HRV-10 at an m.o.i. of 10 (virus particles/cell) was addedto each well along with the appropriate amount of compound of formula Ior II. The plate was then incubated at 34° C. After 3 hours thesupernatant was removed, and 200 μL of media was added along with thesame concentration of compound as used in the beginning of the assay.The plates were incubated for 3-4 days at 34° C. To determine the amountof cell growth, an MTT assay (0.5 mgs/mL), as described in Mosmann, T.J. J. Immunol. Methods 1983, 65, 55-63, which is incorporated herein byreference, was performed on the cells, and the plate was read at anoptical density of 540 nm. The results of the assay are set forth inTable 3. The compounds were tested against control compound Pirodavir,obtained from Janssen Pharmaceuticals. The EC₅₀ was measured asdescribed above for the HI-HeLa Cell Culture Assay.

TABLE 3 Compound # ED₅₀ (μM)  3 0.04  4 0.15  5 0.001 11 0.0007 12 0.00413 0.0004 27 0.07 85 0.005 pirodavir 0.0075

Anticoxsackieviral HI-HeLa Cell Culture Assay

The ability of compounds to protect calls against CVB-3 infection wasmeasured by the XTT dye reduction method, which is described in Weislow,O. S., R. Kiser, D. L. Fine, J. Bader, R. H. Shoemaker, and M. R. Boyd,1989, J. Natl. Cancer Inst. 81:577-586, which is incorporated herein byreference. Specifically, HI-HeLa cells were infected wtih CVD-3 at amultiplicity of infection (m.o.i.) of 0.08 or mock-infected with mediumonly. Infected or mock-infected cells were resuspended at 8×10⁵ cellsper mL and incubated with appropriate concentrations of compound. Oneday later, XTT/PMS were added to test plates and the amount of formazanproduced was quantified spectrophotometrically at 450/650 nm. The EC₅₀was calculated as the concentration of compound that increased thepercentage of formazan production in compound-treated, virus-infectedcells to 50% of that produced by compound free, mock-infected cells. The50% cytotoxic dose (CC₅₀) was calculated as the concentration of drugthat decreased the percentage of formazan produced in compound treated,mock-infected cells to 50% of that produced in compound-free,mock-infected cells. The therapeutic index (TI) was calculated bydividing the CC₅₀ by the EC₅₀.

The Coxsackie strain B-3 (CVB-3) was purchased from American TypeCulture Collection (ATCC). Virus stocks were propgated and antiviralassays were performed in Hi-HeLa cells (ATCC). Cells were grown inMinimal Essential Medium with 10% fetal bovine serum.

The compunds were tested against control compound WIN 54954, obtainedfrom Sterling Winthrop Pharmaceuticals, and control compound Pirodavir,obtained from Janssen Pharmaceuticals.

TABLE 4 Compound # EC₅₀ (μM) CC₅₀ (μM) TI  3 39.8 >320 >8 118.9 >320 >35 13 >100 >100 21 158 >320 >2 23 >100 >100 24 10 10 1 27 20102.7 >5 37 17.8 >100 >5.6 41 >100 >100 WIN 54954 >100 >100Pirodavir >100 >100

We claim:
 1. A compound of the formula (I):

wherein R₁ is H, F, an alkyl group, OH, SH, an O-alkyl group, or anS-alkyl group; R₂ and R₅ are independently selected from H,

or an alkyl group, wherein said alkyl group is different from

with the proviso that at least one of R₂ or R₅ must be

and wherein, when R₂ or R₅ is

X is ═CH or ═CF and Y₁ is ═CH or ═CF or X and Y₁ together with Q′ form athree-membered ring in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is —CH— or—CF—, and Y₁ is —CH—, —CF—, or —C(alkyl—, where R₁₀ and R₁₁independently are H, a halogen, or an alkyl group, or, together with thecarbon atom to which they are attached, form a cycloalkyl group or aheterocycloalkyl group, or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₁ is—O—, —S—, —NR₁₂—, —C(R₁₃)(R₁₄)—, —C(O)—, —C(S)—, or —C(CR₁₃R₁₄)— whereinR₁₂ is H or alkyl, and R₁₃ and R₁₄ independently are H, F, or an alkylgroup, or, together with the atom to which they are bonded, form acycloalkyl group or a heterocycloalkyl group; and A₁ is C, CH, CF, S, P,Se, N, NR₁₅, S(O), Se(O), P—OR₁₅, or P—NR₁₅R₁₆ wherein R₁₅ and R₁₆independently are an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group, or, together with the atomto which they are bonded, form a heterocycloalkyl group; and D₁ is amoiety with a lone pair of electrons capable of forming a hydrogen bond;and B₁ is H, F, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈,—NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈ wherein R₁₇, R₁₈, and R₁₉ independently areH, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, or an acyl group, or, wherein any two of R₁₇,R₁₈, and R₁₉, together with the atom(s) to which they are bonded, form aheterocycloalkyl group; and with the provisos that when D₁ is the moiety≡N with a lone pair of electrons capable of forming a hydrogen bond, B₁does not exist; and when A₁ is an sp³ carbon, B₁ is not —NR₁₇R₁₈ when D₁is the moiety —NR₂₅R₂₆ with a lone pair of electrons capable of forminga hydrogen bond, wherein R₂₅ and R₂₆ are independently H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group; and wherein D₁—A₁—B₁ optionally forms a nitro groupwhere A₁ is N; and wherein, when R₂ or R₅ is

X is ═CH or ═CF and Y₂ is ═C, ═CH or ═CF, or X and Y₂ together with Q′form a three-membered ring in which Q′ is —C(R₁₀)(R₁₁)— or —O—, X is—CH— or —CF—, and Y₂ is —CH—, —CF—, or —C(alkyl)—, where R₁₀ and R₁₁independently are H, a halogen, or an alkyl group, or, together with thecarbon atom to which they are attached, form a cycloalkyl group or aheterocycloalkyl group, or X is —CH₂—, —CF₂—, —CHF—, or —S—, and Y₂ is—O—, —S—, —N(R′₁₂)—, —C(R′₁₃)(R′₁₄)—, —C(O)—, —C(S)—, or —C(CR′₁₃R′₁₄)—wherein R′₁₂ is H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —OR′₁₃,—NR′₁₃R′₁₄, —C(O)—R′₁₃, —SO₂R′₁₃, or —C(S)R′₁₃, and R′₁₃ and R′₁₄,independently are H, F, or an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, or a heteroaryl group or,together with the atom to which they are attached, form a cycloalkylgroup or a heterocycloalkyl group; and wherein any combination of Y₂,A₂, B₂, and D₂ forms a cycloalkyl group, a heterocycloalkyl group, anaryl group, or a heteroaryl group; and A₂ is C, CH, CF, S, P, Se, N,NR₁₅, S(O), Se(O), P—OR₁₅, or P—NR₁₅R₁₆ wherein R₁₅ and R₁₆independently are an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, or a heteroaryl group or, together with the atomto which they are bonded, form a heterocycloalkyl group; and D₂ is amoiety with a lone pair of electrons capable of forming a hydrogen bond;and B₂ is H, F, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, —OR₁₇, —SR₁₇, —NR₁₇R₁₈,—NR₁₉NR₁₇R₁₈, or —NR₁₇OR₁₈ wherein R₁₇, R₁₈, and R₁₉ independently areH, an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, or an acyl group, or, wherein any two of R₁₇,R₁₈, and R₁₉, together with the atom(s) to which they are bonded, form aheterocycloalkyl group; R₃ and R₆ are independently H, F, or an alkylgroup; R₄ is H, OH, or a suitable organic moiety; Z and Z₁ areindependently H, F, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂,—SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃), —PO(NR₂₁R₂₂)(NR₂₃R₂₄),—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃, wherein R₂₁, R₂₂, R₂₃, and R₂₄are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or acylgroup, or a thioacyl group, or wherein any two of R₂₁, R₂₂, R₂₃, andR₂₄, together with the atom(s) to which they are bonded, form aheterocycloalkyl group; or Z₁, as defined above, together with R₁, asdefined above, and the atoms to which Z₁ and R₁ are bonded, form acycloalkyl or heterocycloalkyl group, or Z and Z₁, both as definedabove, together with the atoms to which they are bonded, form acycloalkyl or heterocycloalkyl group; or a pharmaceutically acceptableprodrug, salt, or solvate thereof; and wherein said compound,pharmaceutically acceptable prodrug, salt, or solvate thereof, hasantipicornaviral activity with an EC₅₀ less than or equal to 100 μM inthe HI—HeLa cell culture assay.
 2. A compound of claim 1 wherein R₁ is Hor F, or a pharmaceutically acceptable prodrug, salt, or solvatethereof.
 3. A compound of claim 1 wherein R₄ is an acyl group or asulfonyl group, or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 4. A compound of claim 1, wherein at least one of R₂ orR₅ is

or a pharmaceutically acceptable prodrug, salt, or solvate thereof.
 5. Acompound according to claim 4, wherein D₁ is —OR₂₅, ═O, ═S, ≡N, ═NR₂₅,or —NR₂₅R₂₆, wherein R₂₅ and R₂₆ are independently H, an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group, or, together with the nitrogen atom to which they arebonded, form a heterocycloalkyl group; or a pharmaceutically acceptableprodrug, salt, or solvate thereof.
 6. A compound according to claim 5,wherein D₁ is ═O; or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 7. A compound according to claim 4, wherein A₁ is C,CH, S, or S(O); or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 8. A compound according to claim 7 wherein A₁ is C; ora pharmaceutically acceptable prodrug, salt, or solvate thereof.
 9. Acompound according to claim 4 wherein B₁ is NR₁₇R₁₈, wherein R₁₇ and R₁₈are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup, or wherein R₁₇ and R₁₈, together with the atom(s) to which theyare bonded, form a heterocycloalkyl group; or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof.
 10. A compound accordingto claim 1, wherein at least one of R₂ or R₅ is

or a pharmaceutically acceptable prodrug, salt, or solvate thereof. 11.A compound according to claim 10, wherein D₂ is —OR₂₅, ═O, ═S, ≡N,═NR₂₅, or —NR₂₅R₂₆, wherein R₂₅ and R₂₆ are independently H, an alkylgroup, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or aheteroaryl group, or, together with the atom to which they are bonded,form a heterocycloalkyl group; or a pharmaceutically acceptable prodrug,salt, or solvate thereof.
 12. A compound according to claim 11, whereinD₂ is═O; or a pharmaceutically acceptable prodrug, salt, or solvatethereof.
 13. A compound according to claim 10, wherein A₂ is C, CH, S,or S(O); or a pharmaceutically acceptable prodrug, salt, or solvatethereof.
 14. A compound according to claim 13, wherein A₂ is C; or apharmaceutically acceptable prodrug, salt, or solvate thereof.
 15. Acompound according to claim 10 wherein B₂ is NR₁₇R₁₈, wherein R₁₇ andR₁₈ are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup, or wherein R₁₇ and R₁₈, together with the atom(s) to which theyare bonded, form a heterocycloalkyl group; or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof.
 16. A compound accordingto claim 1, wherein A₁ is C, CH, S, or S(O) and wherein A₂ is C, CH, S,or S(O); or a pharmaceutically acceptable prodrug, salt, or solvatethereof.
 17. A compound according to claim 1 wherein Z and Z₁ areindependently H, an aryl group, or a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂,—SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃; wherein R₂₁, R₂₂, and R₂₃ areindependently H, an alkyl group, a cycloalkyl group, a heterocycloalkylgroup, an aryl group, a heteroaryl group, an acyl group, or a thioacylgroup, or wherein any two of R₂₁, R₂₂, and R₂₃, together with theatom(s) to which they are bonded, form a heterocycloalkyl group, or Zand Z₁, together with the atoms to which they are bonded, form aheterocycloalkyl group; or a pharmaceutically acceptable prodrug, salt,or solvate thereof.
 18. A compound according to claim 1, wherein saidcompound has the formula II:

wherein R₃₁ is H, F or an alkyl group; R₃₂ is selected from one of thefollowing moieties:

wherein R₃₅ is H, an alkyl group, an aryl group, —OR₃₈, or —NR₃₈R₃₉, andR₃₆ is H or an alkyl group, or R₃₅ and R₃₆, together with the atom(s) towhich they are attached, form a heterocycloalkyl group or a heteroarylgroup; R₄₁ is H, an alkyl group, an aryl group, —OR₃₈, —SR₃₉, —NR₃₈R₃₉,—NR₄₀NR₃₈R₃₉, or —NR₃₈OR₃₉, or R₄₁ and R₃₆, together with the atom towhich they are attached, form a heterocycloalkyl group; and R₃₇ is analkyl group, an aryl group, or —NR₃₈R₃₉; wherein R₃₈, R₃₉, and R₄₀independently are H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, or an acylgroup, or, wherein any two of R₃₈, R₃₉, and R₄₀, together with theatoms(s) to which they are bonded, form a heterocycloalkyl group, n is0, 1 or 2; R₃₃ is H or an alkyl group; R₃₄ is an alkyl group, acycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroarylgroup, an O-alkyl, an O-cycloalkyl group, an O-heterocycloalkyl group,an O-aryl group, an O-heteroaryl group, an S-alkyl group, an NH-alkylgroup, and NH-aryl group, an N,N-dialkyl group, or an N,N-diaryl group;and Z and Z₁ are independently H, F, an alkyl group, a cycloalkyl group,a heterocycloalkyl group, an aryl group, a heteroaryl group, —C(O)R₂₁,—CO₂R₂₁, —CN, —C(O)NR₂₁,R₂₂, —C(O)NR₂₁OR₂₂, —C(S)R₂₁, —C(S)NR₂₁R₂₂,—NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂, —SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁,—PO(OR₂₁)₂, —PO(R₂₁)(R₂₂), —PO(NR₂₁R₂₂)(OR₂₃), —PO(NR₂₁R₂₂)(NR₂₃R₂₄),—C(O)NR₂₁NR₂₂R₂₃, or —C(S)NR₂₁NR₂₂R₂₃, wherein R₂₁, R₂₂, R₂₃, and R₂₄are independently H, an alkyl group, a cycloalkyl group, aheterocycloalkyl group, an aryl group, a heteroaryl group, an acylgroup, or a thioacyl group, or wherein any two of R₂₁, R₂₂, R₂₃, andR₂₄, together with the atom(s) to which they are bonded, form aheterocycloalkyl group, or Z and Z₁, together with the atoms to whichthey are bonded, form a heterocycloalkyl group; or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof.
 19. A compound accordingto claim 18 wherein Z and Z₁ are independently H, an aryl group, or aheteroaryl group, —C(O)R₂₁, —CO₂R₂₁, —CN, —C(O)NR₂₁R₂₂, —C(O)NR₂₁OR₂₂,—C(S)R₂₁, —C(S)NR₂₁R₂₂, —NO₂, —SOR₂₁, —SO₂R₂₁, —SO₂NR₂₁R₂₂,—SO(NR₂₁)(OR₂₂), —SONR₂₁, —SO₃R₂₁, —C(O)NR₂₁NR₂₂R₂₃, or—C(S)NR₂₁NR₂₂R₂₃; wherein R₂₁, R₂₂, and R₂₃ are independently H, analkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, or an acyl group, or wherein any two of R₂₁,R₂₂, and R₂₃, together with the atom(s) to which they are bonded, form aheterocycloalkyl group, or Z and Z₁, together with the atoms to whichthey are bonded, form a heterocycloalkyl group; or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof.
 20. A compound accordingto claim 18 wherein R₃₂ is selected from one of the following moieties:

wherein R₃₅ is H, an alkyl group, an aryl group, —OR₃₈, —SR₃₉, —NR₃₈R₃₉,—NR₄₀NR₃₈R₃₉, or —NR₃₈OR₃₉, and R₃₆ is H or an alkyl group, or R₃₅ andR₃₆, together with the atom to which they are attached, form aheterocycloalkyl group or a heteroaryl group; R₃₇ is an alkyl group, anaryl group, or —NR₃₈R₃₉; wherein R₃₈, R₃₉, and R₄₀ independently are H,an alkyl group, a cycloalkyl group, a heterocycloalkyl group, an arylgroup, a heteroaryl group, or an acyl group, or, wherein any two of R₃₈,R₃₉, and R₄₀, together with the atom(s) to which they are bonded, form aheterocycloalkyl group, n is 0, 1 or 2; or a pharmaceutically acceptableprodrug, salt, or solvate thereof.
 21. A compound according to claim 1,wherein said compound has the formula III:

wherein R₂ is CH₂CH₂C(O)NHCPh₃, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃, R₂is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃, R₂ isCH₂NHC(O)CH₃; R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃, R₁ is H, Z is H, andZ₁ is CO₂CH₂CH₃, and R₂ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is CO₂CH₃, and Z₁ is H, R₂ isCH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₃, R₂ is CH₂CH₂S(O)CH₃,R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z isH, and Z₁ is C(O)CH₃, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ isCN, R₂ is CH₂NHC(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃, R₂ isCH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH(CH₃)₂, R₂ isCH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is C(O)N(CH₃)₂, R₂ isCH₂CH₂C(O)NH₂; R₁ is H, Z is H, and Z₁ is C(O)Ph, R₂ is CH₂CH₂C(O)NH₂;R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CH₂Cl, or R₂ isCH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is

or a pharmaceutically acceptable prodrug, salt, or solvate thereof. 22.A compound according to claim 1, wherein said compound has the formulaIV:

wherein R₂ is CH₂CH₂C(O)NH₂, X₁ is H, F, or Cl, and X₂ is H, F, or Cl;or a pharmaceutically acceptable prodrug, salt, or solvate thereof. 23.A compound according to claim 22 wherein X₁ is Cl and X₂ is H; X₁ is Fand X₂ is H; or X₁ is H and X₂ is F; or a pharmaceutically acceptableprodrug, salt, or solvate thereof.
 24. A compound according to claim 1,wherein said compound has the formula V:

wherein: R₄ is PhCH₂OC(O), X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z isH, and Z₁ is CO₂CH₂CH₃; or R₄ is CH₃CH₂CH₂SO₂, X₁ is H, R₂ isCH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃; or R₄ is PhCH₂SO₂,X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is CO₂CH₂CH₃; orR₄ is CH₃CH₂SO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁is CO₂CH₂CH₃; or R₄ is PhSO₂, X₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Zis H, and Z₁ is CO₂CH₂CH₃; or a pharmaceutically acceptable prodrug,salt, or solvate thereof.
 25. A compound according to claim 1, whereinsaid compound has the formula VI or VII:

or a pharmaceutically acceptable prodrug, salt, or solvate thereof. 26.A compound according to claim 1, wherein said compound has the formulaVIII:

wherein X₁ is F, R₂ is CH₂CH₂C(O)NH₂, Y is CH, Z is H, and Z₁ isCO₂CH₂CH₃; or X₁ is H, R₂ is CH₂CH₂C(O)NH₂, Y is N, Z is H, and Z₁ isCO₂CH₂CH₃, or X₁ is H, R₂ is CH₂CH₂C(O)NH₂, Y is CH, Z is H, and Z₁ isC(O)N(CH₃)OCH₃; or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 27. A compound according to claim 1, wherein saidcompound has the formula III:

wherein R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is CH₃ and Z₁ is CO₂CH₂CH₃. R₂is CH₂CH₂C(O)NH₂, R₁ is H, and Z and Z₁ together form

or wherein R₂ is CH₂CH₂C(O)NH₂, R₁ is H, Z is H, and Z₁ is selectedfrom:

or a pharmaceutically acceptable prodrug, salt, or solvate thereof. 28.A compound according to claim 1, wherein said compound has the formulaXIV:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(O)NH₂, Z is H, Z₁ is CO₂CH₂CH₃and R₃ is CH₂Ph and R₄ is

R₃ is

and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is

and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is

and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂CH₃ and R₄ is

R₃ is CH₃ and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is

and R₄ is

R₃ is

and R₄ is

R₃ is

and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

R₃ is CH₂Ph and R₄ is

or R₃ is CH₂Ph and R₄ is

or a pharmaceutically acceptable prodrug, salt or solvate thereof.
 29. Acompound according to claim 1, wherein said compound has the formulaXIV:

wherein R₆ is H, R₁ is H, R₃ is CH₂Ph, R₂ is CH₂CH₂C(O)NH₂, Z and Z₁together form

and R₄ is

or a pharmaceutically acceptable prodrug, salt or solvate thereof.
 30. Acompound according to claim 1, wherein said compound has the formulaXIV:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(O)NH₂, R₄ is

and R₃ is

Z is H and Z₁ is CO₂CH₂CH₃ R₃ is CH(OH)CH₃, Z is H and Z₁ is CO₂CH₂CH₃R₃ is

Z is H and Z₁ is CO₂CH₂CH₃ R₃ is CH₂Ph, Z is H and Z₁ is C(O)N(OH)CH₃ R₃is

Z is H and Z₁ is CO₂CH₂CH₃ R₃ is

Z is H and Z₁ is CO₂CH₂CH₃ R₃ is CH₂CH(CH₃)₂, Z is H and Z₁ is CO₂CH₂CH₃R₃ is CH₂SCH₃, Z is H and Z₁ is CO₂CH₂CH₃ or R₃ is CH₂SCH₂CH₃, Z is Hand Z₁ is CO₂CH₂CH₃ or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 31. A compound according to claim 1, wherein saidcompound has the formula IX:

wherein R₆ is H, R₁ is H, R₂ is CH₂CH₂C(C(O)NH₂, Z is H, and Z₁ isCO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

or Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

or a pharmaceutically acceptable prodrug, salt or solvate thereof.
 32. Acompound according to claim 1, wherein said compound has the formula IX:

wherein R₆ is H, R₃ is CH₂Ph, R₂ is CH₂CH₂C(O)NH₂, and R₁ is OH, Z is H,Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is

and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂C(CH₃)₃, and R₄ is

R₁ is H, Z and Z₁ together form

and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃ and R₄ is

R₁ is H, Z is CH₃, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z and Z₁ together form

and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is CH₃, Z₁ is CO₂CH₂CH₃, and R₄ is

R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

or R₁ is H, Z is H, Z₁ is CO₂CH₂CH₃, and R₄ is

or a pharmaceutically acceptable prodrug, salt or solvate thereof.
 33. Acompound according to claim 1, wherein said compound has the formula IX:

wherein R₆ is H, R₂ is CH₂CH₂C(O)NH₂, R₁ is H, and Z is H, Z₁ isCO₂CH₂CH₃, R₃ is

and R₄ is

Z is CH₃, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Z is CH₃, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z and Z₁ together form

R₃ is

and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is CH₂Ph, and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

or Z is H, Z₁ is CO₂CH₂CH₃, R₃ is

and R₄ is

or a pharmaceutically acceptable prodrug, salt or solvate thereof.
 34. Apharmaceutical composition comprising: (a) a therapeutically effectiveamount of a compound as defined in claim 1 or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof; and (b) a pharmaceuticallyacceptable carrier, diluent, vehicle, or excipient.
 35. A method oftreating a mammalian disease condition mediated by picornaviral proteaseactivity that comprises administering to a mammal for the purpose ofsaid treating a therapeutically effective amount of a compound asdefined in claim 1 or a pharmaceutically acceptable prodrug, salt, orsolvate thereof.
 36. A method of inhibiting the activity of apicornaviral 3C protease that comprises contacting the picornaviral 3Cprotease for the purpose of said inhibiting with an effective amount ofa compound as defined in claim 1 or a pharmaceutically acceptableprodrug, salt, or solvate thereof.
 37. A method of inhibiting theactivity of a rhinoviral protease that comprises contacting therhinoviral protease for the purpose of said inhibiting with an effectiveamount of a compound as defined in claim 1 or a pharmaceuticallyacceptable prodrug, salt, or solvate thereof.
 38. A method of making acompound according to claim 1, comprising converting a compound offormula Q

wherein R₁, R₂ and R₅ are as defined in claim 1, and P₁ is a protectivegroup, or a salt or solvate thereof, to a compound of formula I, asdefined in claim 1, or a pharmaceutically acceptable prodrug, salt orsolvate thereof.
 39. A method according to claim 38, wherein P₁ isbenzyloxy carbonyl or t-butoxycarbonyl.
 40. A method a making a compoundaccording to claim 1, comprising converting a compound of the formula B:

wherein R₁, R₂ and R₅ are as defined in claim 1, or a salt or solvatethereof, to a compound of formula I, as defined in claim 1, or apharmaceutically acceptable prodrug, salt or solvate thereof.
 41. Amethod of making a compound according to claim 1, comprising convertinga compound of formula O,

wherein R₁, R₂, R₅, Z and Z₁ are as defined in claim 1 and P₁ is aprotective group, or a salt or solvate thereof, to a compound of formulaI, as defined in claim 1, or a pharmaceutically acceptable prodrug, saltor solvate thereof.
 42. A method according to claim 41, wherein P₁ isbenzyloxy carbonyl or t-butoxycarbonyl.
 43. A method of preparing acompound according to claim 1, comprising converting a compound offormula P:

wherein R₁, R₂, R₅, Z and Z₁ are as defined in claim 1, or a salt orsolvate thereof, to a compound of formula I, as defined in claim 1, or apharmaceutically acceptable prodrug, salt or solvate thereof.
 44. Acompound according to claim 1, or a pharmaceutically acceptable prodrugor a pharmaceutically acceptable salt, solvate, or any crystal formthereof, wherein said antipicornaviral activity is antirhinoviralactivity.
 45. A compound according to claim 1, or a pharmaceuticallyacceptable prodrug or a pharmaceutically acceptable salt, solvate, orany crystal form thereof, wherein said antipicornaviral activity isanticoxsackieviral activity.